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1. CAN I USE QX CAPSULES FOR NON ADHERENT CELLS Yes Cells in suspension such as blood cells bacteria and protozoa can be attached to the capsule membrane coated with attachment factor such as poly L lysine or gelatin The cells can be attached to the coated membrane by incubating or by centrifuging For detailed protocols see the QX 02 User Manual ARE THERE SPECIFIC STAINS FOR DIFFERENT CELLULAR COMPARTMENTS Since the imaging contrast in WETSEM technology is based on variations in atomic numbers heavy metal stains give the best contrast when imaging biological samples Due to different affinities of heavy metals to various molecules some cellular structures stain more strongly and can be visualized For example osmium tetroxide has a high affinity for lipids and can be used for staining lipid vesicles Most of the heavy metals stain the nuclei and some of them such as gold chloride can be used for staining chromosomes However since the heavy metal stains are usually quite non specific for detailed localization studies immunogold labeling may be required CAN I CARRY OUT INTRACELLULAR IMMUNOLABELING WITH WETSEM TECHNOLOGY Yes Colloidal gold particles are well visualized with WETSEM technology and intracellular antigens can be labeled on fixed permeabilized cells using commercially available gold conjugates Generally smaller gold particles less than 10 nm are better suited for intracellular labeling Parti
2. technology leads to a better understanding of biomedical and metabolic processes for more effective medical chemical biological and industrial research An ultra thin membrane completely isolates hydrated samples from the microscope chamber s vacuum This unique receptacle permits electron microscopy of samples held in a liquid medium By combining the functionality of a cell culture dish or tissue specimen holder with an electron transparent vacuum tight window OX capsules enable high resolution imaging of fully hydrated samples of human animal and plant cells tissues and fluids microbial cells and other types of fluids A new approach to cell and tissue imaging and analysis Eliminate timely sample preparation Attain EM level resolution for all types of hydrated samples 16 Electron Microscopy Sciences gt 1560 Industry Road gt PO Box 550 gt Hatfield PA 19440 gt Wa GO ee SE Cross section of mouse Porcine skeletal muscle Arabidopsis thaliana stigma spinal cord uranyl acetate v uranyl acetate v osmium tetroxide v Mitochondria gold immunolabeled Trypanosoma brucei NIH 3T3 cell in HeLa cells v uranyl acetate v osmium tetroxide v p 4 F E 1 2 Epidermal growth factor receptors immuno labeled with 40 nm gold nanoparticles on A431 cells imaged at 30 kv The distribution of the receptors on the cell membrane is shown by counterstaining with uranyl acetate In collaboration with Professor Joseph
3. Graphic User Guide WETSEM Imaging Quick Guide SK 102 302 gt The SK 102 302 Starter Kit includes QX 102 Capsules and QX 302 Capsules and all the necessary accessories to get started with the QuantomiX WETSEM technology for all types of samples The kit contains QX 102 Capsules 12 units QX 302 Capsules 6 units MP 10 Multi well Plates 2 units MP 12 Multi capsule Plates 2 units MA 4 Multi well Aspirator QX Imaging Buffer QX 302 Imaging Buffer QX 102 Calibration Capsule User Manuals Applications Manual Instructional Training Movie Graphic User Guide WETSEM Imaging Quick Guide 0 ON DW KR WD PI 24 Particle Imaging Kit gt The kit contains Aspirator Tips 24 units 2 Poly L Lysine 1 WT Iml 3 Poly sodium 4 styrenesulfonate 0 3 WT I ml 4 Distilled water 4ml Me FREQUENTLY ASKED T QUESTIONS ABOUT WETSEM TECHNOLOGY This section covers the most frequently asked questions Topics gt SEM amp IMAGING gt APPLICATIONS gt HANDLING THE CAPSULES gt SAMPLE TREATMENT IN QX CAPSULES gt CELL BIOLOGY WITH QX 102 CAPSULES SEM and Imaging v Q WILL THE QX CAPSULES FIT MY SEM A The QX capsules are designed to fit into the specimen stage of most types of SEMs The QX capsule works with SEMs equipped with BSE Back Scattered Electrons detectors Either variable pressure or high vacuum mode can be used Adapters are available for different stub holder dimensio
4. a ie ans ak lt j A i a a _ P i A i _ S F aa Y g a i a f ee gt Pi P e Lipid body visualization in differentiating 3T3 L1 pre adipocyte cells QuantomiX tel 215 412 8400 gt fax 215 412 8450 gt email sgkcck aol com gt web www emsdiasum com 9 Life Science Application Note lt Lipid Research with QX Capsules WETSEM technology is particularly suitable for lipid visualization resulting in high signal to noise ratio tailored for lipid imaging The sample placed in a sealed capsule is separated from the vacuum by a thin electron transparent membrane allowing imaging of wet un embedded cells and tissues The direct immediate and reliable visualization of lipids with QX capsules is a major step forward for lipid research ta ty 1r ehh Th z dv a Intramyocellular lipids in diabetic skeletal muscle in rodent Cross section of a mouse spinal cord The myelin ring unstained In collaboration with Dr N Kaiser Hadassah are shown in black unstained In collaboration wit Medical school Teva Pharmaceuticals Lipid bodies in cultured HeLa cells stained with Lipid bodies in human polymorphonuclear cells Osmium tetroxide of peripheral white blood cells stained with Osmium tetroxide Benefits High resolution imaging of wet samples m Artifact free Minimal sample preparation Optimal signal to noise ratio Quantifiable Lipid visualization QUANTOMIX I0 Ele
5. 102 capsules without damaging the capsule membrane Aspirator tip should be used with a standard calibrated pipette to aspirate liquids from the QX capsule liquid dish To apply liquids into the liquid dish use conventional tips The supplied distilled water is used to wash the membrane from residual reagents 4 QX 302 CAPSULES The QX 302 Capsules are used for imaging various thick non adherent samples such as tissue biopsies plants and material specimens in their natural wet state The capsule is suitable for variable sample sizes with a maximum diameter of 3mm and thickness of up to Imm No coating or embedding of the sample are required enabling electron microscopy imaging with easy sample preparation comparable to light microscopy The QX 302 Capsules are supplied sterile for single use in boxes of 6 ACCESSORIES FOR THE QX 302 CAPSULES MP 12 Multi capsule Plate gt The Multi capsule Plate is designed to enable parallel handling of a number of individual QX 302 Capsules It serves for holding the capsules during specimen preparation and for storage The Multi capsule Plates are supplied sterile and intended for single use QX 302 Imaging Buffer gt QX 302 Imaging Buffer is a solution optimized for imaging samples in an SEM with QX 302 capsules and is formulated to minimize damage to the samples by the electron beam It is applied on samples in the QX 302 capsules prior to SEM imaging The QX 302 Imaging Buffer includes
6. DOES THE ELECTRON BEAM CAUSE DAMAGE TO MY SAMPLE Fluids are very good conductors of heat Therefore the damage by the electron beam is very small The QX 02 Imaging Buffer is specially formulated to minimize the damage caused by the beam and should be used whenever applicable Also to minimize the damage it is recommended to work with lower beam currents i e reduced spot size ARE THERE ANY RESTRICTIONS TO THE ELECTRON BEAM CURRENT There are some considerations to the choice of beam parameters and the recommended parameters can be found in the QX 102 User Manual Generally higher beam currents result in stronger signals but increase the possibility of damage to the sample and affect the resolution Lower beam currents minimize the damage and result in better resolution at high magnifications but may give a signal that is not satisfactory The optimal beam current depends on the sample and is determined empirically IS THERE A PROBLEM OF CHARGING WITH QX CAPSULES No Since the sample inside the QX 102 capsule is wet and the capsule itself is conductive there is no charging ARE THE QX 102 CAPSULES COMPATIBLE WITH LIGHT MICROSCOPES Yes The membrane of the QX 102 capsule as well as the MP 10 multi well plate are transparent to light and allow imaging of samples with a light microscope The imaging should be done only with the QX 102 capsules properly placed in the MP 10 multi well plate which is especially designed fo
7. University School of Medicine New Heaven CT Fig 4 Lipid bodies in human white blood cells After separation cells were plated on the QX Capsule membrane Efficient attachment and spreading of the population of interest were achieved Fixation and consequent uranyl acetate and osmium staining enhanced visualization of cellular organelles and highlighted the lipid bodies Fig 4 White Blood Cells Advantages e High resolution imaging of wet samples e Molecular immunolabeling e Visualization of intra and extracellular structures Minimal sample preparation e Imaging of adherent and non adherent cells e Artifact free QuantomiX 14 Electron Microscopy Sciences gt 1560 Industry Road gt PO Box 550 gt Hatfield PA 19440 ME APPLICATION NOTES 5 QUANTOMLX QuantomiX Lipid Imaging That Counts New opportunities for quantitative analysis in metabolic syndrome research IMAGINE BEING ABLE TO PRODUCE MEANINGFUL IMAGES OF E e a riae TE LIPIDS IN CELLS AND TISSUES THE POSSIBILITIES ARE ENDLESS QUANTOMIX WETSEM TECHNOLOGY ENABLES POWERFUL pre adipocyte call line days in culture IMAGING AND ANALYSIS OF HYDRATED LIPIDS IN CELLS AND TISSUES FOR FAST AND RELIABLE QUANTIFICATION Monitor lipid accumulation in cells By examining hydrated cell samples you can set up cell based assays that mimic metabolic processes that enable reliable mapping of cell differentiation Your lipid accumulation cell assays do
8. capsules open the package in a sterile environment such as laminar flow hood Take out the desired number of capsules close the box and seal the top cover with tape Store the package in a clean dry place tel 215 412 8400 gt fax 215 412 8450 gt email sgkcck aol com gt web www emsdiasum com FAQ Sample Treatment In QX Capsules wv Q CAN I STORE MY SAMPLES IN QX CAPSULES A Yes Samples can be stored in sealed QX capsules and they remain wet The samples can also be stored in open capsules liquid dishes when properly sealed in the MP 10 multi well plate For sealing of liquid dishes and for storage of biological samples please refer to the QX 102 User Manual HOW DO I APPLY LIQUIDS TO THE QX CAPSULES A Liquids and samples are applied onto the liquid dishes of the capsules using standard lab pipettes When treating multiple samples a repetitive dispensing pipette is most convenient It is important to apply the liquid carefully not to touch the capsule membrane with the pipette tips HOW DO I REMOVE LIQUIDS FROM THE QX CAPSULES The liquids are removed using the MA 4 multi well aspirator designed for safe aspiration of liquids from the capsules Other means should not be used since they may lead to rupturing of the capsule membrane For detailed instructions of working with the MA 4 multi well aspirator refer to the instructions accompanying the product See also the QX 102 User Manual for recommendations for p
9. gt Hatfield PA 19440 ME APPLICATION NOTES 4 Life Science Application Note lt Imaging Wet Cells Bridging The Gap Between Light and Electron Microscopy The need to dehydrate and prepare samples before imaging in an electron microscope has long been a serious limitation for cell biology research Biological specimens are hydrated in their natural state but current methods of imaging these specimens under high resolution require dehydration and extensive preparation On the other hand samples can be viewed in their native state in light microscopes but those are limited to low resolution viewing QuantomiX proprietary WETSEM technology brings together the immediate viewing capabilities of light microscopes and the high resolution capacity of electron microscopy and provides an ideal solution for high resolution imaging of fully wet cells Viewing Wet Samples in Their Native State with WETSEM Technology The solution offered by WETSEM technology enables high resolution imaging of samples without lengthy preparation and with no dehydration artifacts The QX capsules based on WETSEM technology are used for holding biological samples in the electron microscope The samples are kept in a sealed vacuum resistant capsule during the imaging process This method has been used successfully for many types of biological samples including non Mitochondria visualization in C2C12 cells that were fixed and st
10. gt INTRODUC T O Neree Electron Microscopy Sciences and QuantomiX have joined forces here in The United States to market and sell the breakthrough solutions of QuantomiX s proprietary WETSEM Technology This technology enables scanning electron microscopes SEM to image and analyze wet samples such as cells tissue biopsies foods and ink in their native environment Eliminating the need for time consuming preparation procedures the WETSEM Technology ensures that sample integrity is not compromised by artifacts QuantomiX innovative technology opens new opportunities for application specific tools to improve drug discovery and advances treatment and diagnostic solutions for the medical and pharmaceutical markets My owetsem TECHNO LO GY eee Electron Microscopy EM is a prime tool for high resolution imaging which has been the cornerstone of our understanding of living organisms and our material environment Because EM requires samples to be placed in a vacuum it does not lend itself for use with wet samples In the 50 years since its development this drawback has been a major impediment in the use of EM for biomedical research and for many clinical and industrial applications Light Microscopy on the other hand requires only minimal sample preparation However the resolution of light microscopy is limited to a few hundred nanometers The revolutionary technology of QuantomiX solves the problem of preparing wet sa
11. not have to be modified in any way resulting in unprecedented accuracy EM scanning of hydrated tat cells and tissues for Fast implementation with minimal preparation requirements Reliable reproducible and quantifiable results High resolution imaging image fat in tissues Fat cell size distribution and lipid droplet examination in tissues have never been so clear and easy to do Just apply the sample in the OX capsule and expand the horizons of advanced obesity and diabetes research Base tin tissue with Prof 5 Cinti Italy os een ee Ue Aj LEAL t tel 215 412 8400 gt fax 215 412 8450 gt email sgkcck aol com gt web www emsdiasum com l5 Electron microscopy right from the cell culture dish The QuantomiX OX capsule product line enables direct imaging of fully hydrated samples in conventional scanning electron microscopes SEMs Wet samples can be examined eliminating the need for arduous preparation such as critical point drying coating embedding and sectioning Simply place tissue samples directly in the OX capsule or grow cells inside it to achieve high resolution imaging and analysis 1 Prepare multi well plate for OX capsules 2 Apply sample via conventional pipatting 3 Quick seal the single use OX capsule 4 Place the OX capsule in the SEM chamber A new approach to cell and tissue imaging and analysis QuantomiX WETSEM
12. 1M lt 3 5 negative HCI IM 0 1mM 0 4 positive CAN I IMAGE LIVE CELLS WITH QX 102 CAPSULES Cells can be introduced live into the SEM and short time imaging of live cells may be possible However the radiation absorbed by the cells during image scanning is expected to cause structural changes and to affect the viability of the cells Also the contrast between different constituents of native cells may be too low for high resolution imaging and some staining or labeling may be required Live cells or microorganisms can be imaged either in their growth medium or in PBS CAN I IMAGE LIPID RICH STRUCTURES WITH WETSEM TECHNOLOGY WETSEM technology allows analysis of lipid structures in an unperturbed state The ability to image wet samples with SEM avoids the problem of lipid extraction that occurs due to dehydration with organic solvents during lipid imaging with conventional techniques Many lipid structures are visualized without any enhancement If staining is required Osmium tetroxide is well suited for lipids for detailed protocols see the QX 02 User Manual Q CAN I IMAGE FOAMS WITH WETSEM TECHNOLOGY Yes QX 1 02 capsules can be used for imaging cosmetic food or other foam samples Q I WOULD LIKE TO IMAGE UNTREATED FOOD SAMPLES WHAT DETAILS CAN I SEE WITH QX 102 CAPSULES The imaging contrast in QX 02 capsules is created from variations in atomic numbers of sample constituents Thus constituents and
13. K appear perfectly round after the correction see Figure 4 06 gt Set the spot size at 35 of the spot size range 18 gt Obtain a slow scan image and save 07 gt See Table for a working distance suitable for your SEM configuration SAMPLE IMAGING 08 gt Initially use the SE detector and focus on the metal support grid see Figure 1 09 gt Once focused change to the BSE detector OI gt Apply steps l 12 in the above procedure to your sample 10 gt It is recommended to start sample imaging with a slow scan speed 02 gt Adjust contrast and brightness until features of interest are visible a few seconds per frame 03 gt If no features are visible gradually increase spot size I gt Increase contrast drastically The grid should give a strong white signal see Figure 2 04 gt Once features are identified focus adjust contrast and brightness and obtain the 12 gt Move from the grid to one of the windows required images 13 gt Increase magnification until the 500nm silica beads are visible see Figure 3 05 gt Zoom in as required adjusting focus and correcting astigmatism tel 215 412 8400 gt fax 215 412 8450 gt email sgkcck aol com gt web www emsdiasum com l9 gt PRICING E ola its oVe eve e 6 lesa Ce o 6 nian ovsrsievsloveve sin atateielcieie ei e siets le E E ein oie etsioie sie era a aiets r Susie e s oi6 slsieie eieiejeieiniavoiaieieye E oie aise eieieva eio eiaie eloveinioisin o e
14. Schlessinger Department of Pharmacology Yale University School of Medicine 3 3 4 Cross section of rabbit aorta counterstained with uranyl acetate Elastin in the tissue is seen in black imaged at 30 kv In collabora tion with Professor Elazar Edelman Harvard MIT Biomedical Engineering Center 5 gt Ruffled borders in macrophage A 21 cells counterstained with uranyl acetate imaged at 20 kv In collaboration with Professor Paul Matsudaira Whitehead Institute For Biomedical Research MIT tel 215 412 8400 gt fax 215 412 8450 gt email sgkcck aol com gt web www emsdiasum com 17 wire a A B Hematoxylin and eosin stain of normal colon A and IBD B x200 C D Using wet tissue for SEM C en face SEM photograph of normal colon D en face SEM photographs of colon affected by Crohns Disease show marked irregularity of the cell borders with elongation of the crypts orifices x3200 SKIN WITH MINERALIZED ELASTIC FIBERS Pseudoxanthoma elasticum Unfixed and unstained sample Fed and stained sample C collagen Arrows mineralized elastin Electron Microscopy Sciences 1560 Industry Road P O Box 550 gt Hatfield PA 19440 a QUICK USE GUIDE FOR IMAGING ort IMPORTANT NOTES WETSEM images can be obtained with various detectors With most SEM configurations best images are obtained with a BSE detector positioned at the end of the SEM column located un
15. ained with 2 PTA adherent cells and has enabled the collection Mitochondria are easily identified Their pleomorphic of valuable information forms and structural variations are clearly seen Cumont tel 215 412 8400 gt fax 215 412 8450 gt email sgkcck aol com gt web www emsdiasum com l3 Life Science Application Note lt QX Capsules Empowering Imaging Capabilities Fig 1 HeLa cells cultured in the QX capsule in growth medium were fixed with paraformaldehyde and stained with uranyl acetate Cell cell contact between two neighboring cells as well as fillopodia are clearly visible Clear marking of fine intracellular cytoskeletal structures is also seen Fig 2 Wet mast cells were fixed with 4 paraformaldhyde and 0 125 glutaraldehyde and stained with uranyl acetate High resolution imaging with WETSEM enables visualization of individual secretory granules which appear as dark holes Some granules show high electron density Dr Y Satoh Dept of Cell Biology and Functional Morphology Iwate Medical University School of medicine Uchimaro Japan Fig 3 Epidermal growth factor receptor EGFR labeling of A431 cell Anti EGFR antibodies were visualized with 40nm colloidal gold particles Note the very precise visualization of individual gold beads on the cell surface This application is suitable for a variety of intracellular and extracellular immunogold labeling Dr J Schlessinger Dept of Pharmacology Yale
16. cells tissues gt Compositional analysis of wet samples by X ray microanalysis v Wide spectrum of staining and labeling capabilities for cells and tissues Ability to image unstained or unfixed cells and tissues Imaging of both adherent and non adherent cells High resolution histopathology Intracellular imaging in a scanning EM Imaging the entire cell surface Excellent preservation and imaging of lipid structures Easy to automate sample processing and imaging Ability to work with a variety of sample consistencies pastes foam creams emulsions etc v v v v v v v v gt FEATURES VONE ETEEN E E eens ieee gt Direct imaging of all types of wet samples including suspensions emulsions creams cells and tissues Rapid and simplified sample preparation Compositional analysis of wet samples by X Ray analysis Excellent preservation and imaging of lipid structures Easy to automate sample processing and imaging v v v v W Utilizes SEM backscattered electron imaging based on atomic number difference Compatible with light microscopy for comparative studies gt Single use vW gt APPLICATIONS ee eee Lipid Imaging and Analysis Imaging lipid bodies in fully wet cells and tissues Airborne Particles EDS of wet samples with QX capsules Imaging particles in their fully wet environment Experimental Biology Subcellular organelles cytoskeleton and motility cell contacts recep
17. cles less than 10 nm need to be Silver enhanced for imaging HOW DO I CHOOSE THE FIXATION PROTOCOL FOR IMMUNOGOLD LABELING The fixation protocols generally used in immunocytochemistry are also applicable to WETSEM technology The correct choice optimal concentration and time of fixation depend on the nature of the antigen and the antibody Optimal conditions may be established based on your prior experience with the application or by performing preliminary experiments using immuno fluorescence labeling UANTOMLX 6 Electron Microscopy Sciences gt 1560 Industry Road gt P O Box 550 gt Hatfield PA 19440 ME APPLICATION NOTES 1 Material Science Application Note lt Energy Dispersive X Ray Spectroscopy EDS of Wet Samples With QX Capsules EDS is most often used for qualitative elemental analysis to determine which elements are present and to conclude their relative abundance One of the challenges facing this technique is the ability to apply it on wet samples The QX capsule based on QuantomixX proprietary WETSEM technology provides the solution by enabling qualitative EDS measurement and analysis of wet samples such as creams solutions and other liquid containing specimens In Figure 1 a sample of pesticide is imaged and analyzed by EDS The Pesticide elemental composition of the Selected area particles in the sample is determined analysis by selected area EDS Until now specimens
18. ctron Microscopy Sciences gt 1560 Industry Road gt PO Box 550 gt Hatfield PA 19440 ME APPLICATION NOTES 3 Material Science Application Note lt Imaging Particles in Their Fully Wet Environment One of the most valuable parameters in the study of materials is particle characterization Until today particle imaging in Scanning Electron Microscopes was limited to dry samples only As a result valuable information on particles in their native state was lost due to the drying process Furthermore characterization of particle shape size and dispersion in their native wet environment was not feasible at all WETSEM Technology Revealing new perspectives in Material Science Research WETSEM technology presents a breakthrough in imaging and analysis of particles in their native wet state The QX capsule based on QuantomixX proprietary WETSEM technology enables SEM imaging of particles in suspensions pastes gels emulsions creams and liquids of all kinds The QX capsules eliminate the need for time consuming preparation procedures and ensure that artifacts accumulated as a result of drying processes do not compromise sample integrity WETSEM technology presents a tangible solution for formulation and quality control processes in industrial fields a4 tel 215 412 8400 gt fax 215 412 8450 gt email sgkcck aol com gt web www emsdiasum com ll Material Science Application Note lt Meet
19. der the pole piece High probe currents especially when scanning at high magnifications may damage the QX capsule membrane and sample To prevent damage avoid scanning the same area at high magnifications for prolonged periods See Table for the maximum recommended probe current for your SEM configuration v TABLE I SUITABLE PARAMETER RANGE FOR WETSEM IMAGING PARAMETER RECOMMENDED RANGE er O ee ae eo ae Scintilator BSB 1mm Acceptable 10mm In lens Through the lens all detectors v CALIBRATION CAPSULE USE v FN a Figure a Figure 2 a Figure 2 a Figure 4 For first time imaging always use the Calibration Capsule 14 gt If no beads are visible increase the contrast further 5 gt If no beads are visible yet gradually increase the spot size Note that spot size OI gt Reconstitute the Imaging Butter with Imi of double distilled water correlates to probe current See Table for the maximum probe current that 02 gt Place the Calibration Capsule in the Multi well Plate and remove the sealing stub Should be used for WETSEM with your SEM configuration 03 gt Apply 15l of Imaging Buffer into the Calibration Capsule and seal with the stub l6 gt Once the silica beads are visible improve the focus and zoom in 04 gt Place the hydrated Calibration Capsule in the SEM 7 gt Identify the 40nm gold beads focus and correct astigmatism The gold beads should 05 gt Set the acceleration voltage to 30
20. ements present are C O Na Br Mg Al Si S Cl K Ca Fe and Ti 1 00 2 00 3 00 4 00 5 00 6 00 7 00 8 00 9 00 Curno 8 Electron Microscopy Sciences gt 1560 Industry Road gt P O Box 550 gt Hatfield PA 19440 ME APPLICATION NOTES 2 Life Science Application Note lt Imaging Lipid Bodies in Fully Wet Cells and Tissues High resolution imaging of lipid bodies in biological samples is a known challenge Preparation of tissue samples for microscopy examination typically wipes lipids off samples making observation and interpretation of lipid bodies difficult Lipids in cells are observed only indirectly requiring special labeling This has led QuantomiX to develop the QX capsules based on WETSEM technology designed for immediate and quantifiable high resolution lipid imaging The QX capsules present a new opportunity to overcome the limitations of current methods for a better assessment of lipids in cells and tissues Direct Lipid Visualization with WETSEM Technology The QX capsule based on QuantomixX proprietary WETSEM technology enables direct imaging of lipids in fully hydrated samples using standard scanning electron microscopes In the pictures below lipid bodies are clearly visualized in the cellular cytoplasm of 3T3 L1 pre adipocyte cells Ien S p k N aa ae i i i a all Fa 4 a al D a 4 E ea si lt i i k x d A 4 K b F es l s gt o Ps maS fe
21. f the sample To preserve the structure in vacuum the sample must be dehydrated in a controlled manner The sample in the QX capsule is completely isolated from the vacuum and remains at atmospheric pressure Therefore there is no loss of water and no drying of the sample is needed WHAT IS THE RESOLUTION WITH WETSEM TECHNOLOGY In general the resolution is about the same as dry SEM samples depending on the specific sample and the SEM model used see Thiberge et al PNAS Vol 01 No 10 March 9 2004 3346 3351 and Review of Scientific Instruments Vol 75 No 7 July 2004 HOW DEEP BELOW THE MEMBRANE CAN IMAGING BE PERFORMED The beam penetration depth depends on the acceleration voltage For a sample in a water based medium imaged with a 30 kV beam information is retrieved from a depth of approximately 2 4 micrometer At 10 kV this depth is reduced to a few hundred nanometers By varying the acceleration voltage it is possible to obtain unique 3D information from your sample DOES THE MEMBRANE INTERFERE WITH IMAGING The capsule membrane is very thin and flat It is made of material transparent to electrons with energies above a few keV Thus the membrane induces minimal scattering of the beam and of the back scattered electrons detected by the BSE detector The membrane itself is flat and hence does not interfere with the imaging process In X ray mode the membrane produces a constant background that can be subtracted
22. ing the Needs The QX capsules can be used for dispersion characterization of particles in liquids including degree of aggregation homogeneity as well as shape characterization They also permit imaging of emulsions suspensions and other liquids core shell type materials pastes gels creams foams and powders Research has shown that the capsules can FE he aah a provide the scientist with information regarding i chemical composition by EDS of wet samples Important information required for formulation processes in various industries can also be obtained P oun Dead Sea mud Courtesy of AHAVA By using the QX capsule scientists avoid potential problems of film formation and particle aggregation during the drying process The capsules also function as isolation compartments for imaging of air and humidity sensitive materials Advantages m Quick and easy to use m Requires minimal sample preparation m Enables shape size and dispersion characterization m Enables imaging of particles in nano and micro scales Allows chemical composition identification by EDS of wet samples Silica 500nm and gold 40nm particles The QX line of capsules are already in use in biological research and play a major role in the pharmaceutical food detergent paint and ink industries Phase separation in organic encapsulated materials ee a I2 Electron Microscopy Sciences gt 1560 Industry Road gt PO Box 550
23. lsisieve wis sisi hoveisyaseipie eieieie eiinlsie E E T QX 102 PRODUCTS PART QTY PER PRICE rexiasarerk dt Qx 102 Starter Package wio aspirato 1 isoo T Cen biology and general liquid sample Capsules 24 1400 mwene O OOOO i d s muswenaspirasor o f e Parsee magne Jo as RT 56 QX 102 Calibration Capsule QX 302 PRODUCTS PART QTY PER PRICE a saree Stato Tise caps OOO e i Mul capsulePite o s Toxor magng ser Ys a a QX 302 Calibration Capsule COMBINATION PRODUCTS PART QTY PER PRICE NUMBER DESCRIPTION PACKAGE SK 102 302 QX 102 amp QX 302 Combined Starter Kit 4 ae 20 Electron Microscopy Sciences gt 1560 Industry Road gt PO Box 550 gt Hatfield PA 19440
24. mples for high resolution imaging It closes the resolution gap between conventional electron microscopy and light microscopy and offers the convenient sample preparation of light microscopy For the first time rapid and routine EM imaging of biological samples in a wet environment is now possible without the artifacts normally associated with sample preparation gt CONC EP eeeeeesssssee The New QX capsule completely isolates wet samples from the vacuum in the microscope chamber This makes possible the imaging of fully hydrated samples including food cosmetics ink human animal plant and microbial cells tissues and fluids at resolutions unachievable with light microscopy The QX capsule fits the standard SEM specimen stage The capsule combines the function of a specimen holder cell culture dish or a tissue specimen holder with an electron transparent vacuum tight window This unique receptacle permits electron microscopy of samples held in water or any other liquid medium at atmospheric pressure Imaging of samples in the QX capsule can be accomplished with backscattered electron detection x ray detection or light detection to reveal structure as well as material composition gt ADVAN TAG E S eere gt Artifacts associated with sample preparation techniques are eliminated gt Sample preparation time drastically reduced or completely eliminated gt Direct imaging of wet samples food cosmetics inks
25. nces gt 1560 Industry Road gt P O Box 550 gt Hatfield PA 19440 eeccccce Imaging of capsules must begin with a low probe current small spot size Initial probe current should be no higher than 200 pA If the obtained signal is not sufficiently strong probe current can gradually be increased until the desired image is obtained Probe current should not exceed nA Initial magnification should also be low Magnification should be increased gradually until the desired image is obtained Probe current should not exceed nA POCO E EHH ESEEEHSEEEOHHOTEEEOEHEEEEEHOE EE ETOE TEES OOH EEO O ESOS ET ESESESESESESEEEEES FAQ Applications wv Q A CAN I IMAGE ANY LIQUID WITH THE CAPSULES QX capsules are suitable for imaging various types of wet samples and liquids including cells tissues bacteria emulsions oils food samples cosmetics inks particles in solutions etc For solvents that should be avoided please consult the QX 02 User Manual ARE THE QX CAPSULES COMPATIBLE WITH ACIDIC AND BASIC SOLUTIONS The QX capsules have been validated for compatibility with commonly used organic and inorganic acids and bases 5 hour exposure Results are shown in the following table SOLUTION CONCENTRATION PH COMPATIBILITY NaOH 0 1M 0 1mM 13 10 positive NaOH gt 0 1M gt 13 negative Tris hydroxymethyl aminomethane base 0 05M 10 7 adjusted by HCI positive Acetic Acid lt 0 01M gt 3 5 positive Acetic Acid gt 0 0
26. ns Following is a partial list of SEM models compatible with WETSEM technology QX 102 Capsule dimensions in mm 3 14 18 6 gt FEI XL30ESEM XL30ESEM FEG Quanta series Quanta FEG gt JEOL 5600 5900 6000 series gt ZEISS LEO 1450 1500 Series EVO Supra Ultra gt Hitachi 2600 3000 series 4000 series gt Camscan CS3000 range MV2300 range For suitability of other models and availability of Adapters please contact us Q CAN THE CAPSULES BE USED IN HIGH VACUUM MODE A The QX capsules are compatible with high vacuum as well as low vacuum mode The capsules are designed to withstand a pressure difference of up to atmosphere therefore no restrictions exist on vacuum levels and they can also be used at x 0 Torr tel 215 412 8400 gt fax 215 412 8450 gt email sgkcck aol com gt web www emsdiasum com WILL THE SAMPLE IN A QX CAPSULE REMAIN FULLY WET DURING IMAGING IN A SEM Yes The membrane of the QX capsule is impermeable to water and the capsule completely isolates the sample from the vacuum No drying of the sample occurs during imaging or storage WHY IS IT NECESSARY TO DEHYDRATE THE SAMPLE IN CONVENTIONAL SEM IMAGING AND WHY IS NO DRYING NEEDED WITH WETSEM TECHNOLOGY In traditional SEM the sample is placed directly in high vacuum A wet sample exposed to vacuum will lose its water in an uncontrolled dehydration process which often distorts or destroys the structure o
27. r holding the capsules during various manipulations The plate fits into a microscope stage for standard multi well plates CAN I PERFORM EDS WITH WETSEM TECHNOLOGY Yes QX capsules can be used for EDS energy dispersive spectroscopy analysis of samples using a SEM equipped with an EDS system In fact the QX technology uniquely suits EDS of wet samples Note the presence of the metal grid and polymer membrane will be taken into account in the analysis WHAT IMAGING PARAMETERS SHOULD BE USED INWETSEM TECHNOLOGY SEM imaging with QX capsules differs from standard SEM imaging in some aspects Also the factors that affect imaging vary among applications Recommendation for parameters and guidelines for optimization are found in the QX 102 User Manual Optimization of the imaging conditions is best done using the Calibration Capsule cat no RT 56 CAN THE LIQUID SAMPLE LEAK FROM THE CAPSULE AND CONTAMINATE THE DETECTOR OF MY SEM The membrane of the QX capsule is vacuum tight and impermeable to liquids and gases Thus when the capsule is handled correctly there is no leakage of the sample and no contamination Correct handling relates to both physical handling during sample preparation stages and to the parameters used during imaging All care must be taken to prevent physical contact with the membrane Any contact e g with the tip of the pipette or with the hand can potentially tear the membrane Electron Microscopy Scie
28. roper liquid handling during staining and labeling procedures POPS OOOO OEE EE EHO ES HOES E ETOH ET EE ESOS EEE EEHOE EO ETOET EES OOHE EH EESOS ESTES ODES ESESEEEEEEOS FAQ Cell Biology With QX 102 Capsules wv Q WILL MY CELLS GROW ON THE QX 102 CAPSULE MEMBRANE A The QX 02 capsule is designed to be compatible with growth of cells including established cell lines as well as primary cultures Our recommendation is to coat the membrane with a suitable attachment factor prior to cell seeding CAN CELLS BE GROWN IN THE QX 102 CAPSULES FOR MORE THAN ONE DAY Yes The MP 10 multi well plate holder for the capsules is designed to maintain CO and humidity levels so that the capsules can be used as standard cell culture dishes for long term growth of cells During long incubations ensure that the wells on the sides of the MP 10 plate stay filled with water Also it is recommended to change the growth medium of the cells to fresh medium every 2 days Q WHAT ATTACHMENT FACTOR SHOULD USE FOR MY CELL LINE The QX I02 capsule membrane can be coated with various attachment factors such as Fibronectin Collagen Laminin Gelatin poly L lysine or with a combination of them The factors that provide best attachment and growth will depend on the cell type Fibronectin has been found to support growth of many types of cells and thus in many cases will be the recommended first choice Please consult QX 102 User Manual for coating protocols
29. spacers that are used during sample preparation The QX 302 Imaging Buffer is supplied sterile and lyophilized QX 302 Calibration Capsule gt The ox 302 Calibration Capsule is a QX 302 Capsule that has been specially designed to assist first time users in finding optimal imaging conditions for WETSEM imaging in their SEM The Calibration Capsule contains nanoparticles of two different sizes 40 and 500 nm The particles are easily visualized in an SEM and provide a convenient means to calibrate the parameters for optimal wet imaging conditions Electron Microscopy Sciences gt 1560 Industry Road gt P O Box 550 gt Hatfield PA 19440 ME STARTER KITS FOR WETSEM ssi EAEN ATE A EENEN SK 102 24 gt The SK 102 24 Starter Kit includes QX 102 Capsules and all the necessary accessories to get started with the QuantomiX WETSEM technology The kit contains QX 102 Capsules 24 units MP 10 Multi well Plates 2 units MA 4 Multi well Aspirator QX Imaging Buffer QX 102 Calibration Capsule User Manual Applications Manual Instructional Training Movie Imaging Quick Guide no wu KR WW N SK 302 12 gt The SK 302 12 Starter Kit includes QX 302 Capsules and all the necessary accessories to get started with the QuantomiX WETSEM technology The kit contains QX 302 Capsules 12 units 2 MP 12 Multi capsule Plates 2 units 3 QX 302 Imaging Buffer 4 QX 302 Calibration Capsule 5 User Manual
30. structures that have a significant difference in atomic numbers can be visualized without any enhancement The contrast between water and fat is especially well visualized in QxX technology enabling analysis of fat structure and content in food cosmetics and other samples Q I WANT TO LOOK AT COSMETIC CREAM WHICH HAS A PASTE LIKE STRUCTURE CAN I DO THIS WITH QX 102 CAPSULES A Yes Notice that the field you image is that closest to the capsule membrane Therefore the sample has to be in direct contact with the membrane Q IF HAVE A SAMPLE THAT RELEASES GAS INSIDE THE CAPSULE WILL THE MEMBRANE OF THE CAPSULE WITHSTAND THE PRESSURE A As long as the internal pressure does not increase significantly above atmosphere the membrane will withstand the pressure POCO E EEO EE SEE ETOH EOE EE EOESO TEE ESOS EEESOE TEESE TEES OOHEE TOES OES EESESEEESESEEEEEEOOES FAQ Handling The Capsules Q CAN HEAR NOISE OF LOOSE PARTS INSIDE AN UNOPENED PACKAGE OF CAPSULES DOES THIS MEAN THE CAPSULES HAVE BEEN DAMAGED DURING THE SHIPMENT A No The packaging of the capsules is designed to prevent the capsules from moving around and to protect them from damage during shipment To keep the capsules in a dry environment the package contains two cylinders of desiccants which can cause this noise Q CAN I USE ONLY PART OF THE CAPSULES FROM THE PACKAGE AND STORE THE REMAINING UNUSED CAPSULES A Yes In order to maintain sterility of the
31. such as these could not have been analyzed by EDS in their native form and other wet chemical methods had to be used In Figure 2 the image ang EDS spectrum ola O ar a a a ear es ea gel deodorant are clearly seen The different phases are visualized and the chemical composition is directly determined from the EDS spectrum Deadormi 1 00 2 00 3 00 4 00 5 00 6 00 7 00 8 00 9 00 Gurmat tel 215 412 8400 gt fax 215 412 8450 gt email sgkcck aol com gt web www emsdiasum com 7 lt pe a Te ea te Material Science Application Note EDS with WETSEM Technology Stretching Science to New Extremes Wet EDS provides the opportunity to image and analyze the final product without assuming homogeneity In wet EDS sample natural heterogeneity is retained and adjacent structures can be directly compared see analysis below The QX capsules eliminate the need to dry the sample and ensure that artifacts accumulated as a result of drying processes do not compromise sample integrity WETSEM technology presents a tangible solution for formulation and quality control processes in industrial fields Applications include Cosmetics Inks Paints Ceramics Agriculture m Pastes Food SiKa FeKa a ER 1 00 2 00 3 00 4 00 5 00 6 00 7 00 8 00 9 00 CaKa FeKa FeKb 1 00 2 00 3 00 4 00 5 00 6 00 7 00 8 00 9 00 Full frame and spot analysis of Dead Sea mud courtesy of AHAVA In this case the el
32. tor distribution extracellular matrix tissue analysis v v v v v wT wv Industrial applications food oils dyes pharmaceuticals Pathogen characterization Clinical diagnosis Histopathology cytology oncology Tissue engineering implants and prostheses Quality Control Quality Assurance Life Sciences and Medicine Cultured and primary cells Tissue histology Nerve cells and myelin imaging Microbiology Viruses and Plants v v v v v W gt Environmental and toxicological applications gt Nanotechnology and Bio nanotechnology gt Industrial and R amp D Emulsions Suspensions Foods Personal care goods Cosmetics Inks Electron Microscopy Sciences gt 1560 Industry Road gt P O Box 550 gt Hatfield PA 19440 D WETSEM PRODUCT LISTINGS eee lt QX 102 CAPSULES The QX 102 Capsules are used for imaging various wet materials and biological samples such as liquid samples foods cosmetics oils paints etc particles in solutions adherent and non adherent human and animal cells and microorganisms The samples can be visualized either directly or by following appropriate contrast enhancement staining or labeling procedures The capsule serves as a cell culture dish specifically designed for the SEM No coating or embedding of the sample are required enabling electron microscopy imaging with easy sample preparation comparable to light microscopy The QX 102 Capsules are supplied sterile for single
33. use in boxes of 24 ACCESSORIES FOR THE QX 102 CAPSULES MP 10 Multi well Plate gt The Multi well Plate is designed to enable parallel handling of multiple QX 102 Capsules It serves as a well plate for holding the capsules during various manipulations for culturing cells in QX 102 Capsules and for inspection in an inverted light microscope The Multi well Plate is specially designed to maintain humidity of the samples during incubation and is compatible with standard laboratory equipment The Multi well Plates are supplied sterile for single use in boxes of 2 MA 4 Multi well Aspirator gt The Multi well Aspirator system is designed to safely and conveniently aspirate liquids from the QX 102 capsules and is required for applications that need liquid exchange in capsules during sample preparation or cell culturing The Multi well aspirator drains liquids simultaneously from up to four capsules placed in a row in the MP 10 Multi well Plate QX Aspirator Tips gt The QX Aspirator Tips are used to safely and conveniently aspirate liquids from an individual QX 102 capsule with a pipette The tips fit on any standard pipette and are designed to ensure safe liquid removal In cases where multiple capsules will be used or multiple liquid exchanges are required it is recommended to use the MA 4 Multi well Aspirator which drains liquids simultaneously from multiple capsules QX Imaging Buffer gt QX Imaging Buffer is a sol
34. ution optimized for imaging samples in an SEM with QX 102 capsules and is formulated to minimize damage to the samples by the electron beam It is applied on samples in the QX 102 capsules prior to SEM imaging The QX Imaging Buffer is supplied sterile and lyophilized QX 102 Calibration Capsule gt The QX 102 Calibration Capsule is a QX 102 Capsule that has been specially designed to assist first time users in finding optimal imaging conditions for WETSEM imaging in their SEM The Calibration Capsule contains nanoparticles of two different sizes 40 and 500 nm The particles are easily visualized in an SEM and provide a convenient means to calibrate the parameters for optimal wet imaging conditions PI 24 Particle Imaging Kit gt When imaging particles with WETSEM Technology image quality is dependent on the proximity of the particles to the QX capsule membrane The closer the particles are the higher the resolution Coating the membrane with a suitable charged polymer will attract particles to the membrane and improve the resulting image The particle imaging kit provides the WETSEM Technology user with the tools and reagents necessary to coat the QX capsule membrane Poly L Lysine is a positively charged polymer which will attract negatively charged particles Poly sodium 4 sulfonate PSS is a negatively charged polymer which will attract positively charged particles Aspirator tips are designed to safely aspirate the QX

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