PROTOCOLS RNA Handling
Contents
1. into the cap since the smaller fragments produced by multiple LPC points cannot be catapulted so straight to the center of the cap as areas on membrane can be catapulted 6 4 Capture check looking into the cap to see the catapulted samples To control the efficiency of catapulting it is possible to have a look into the collection device e g microfuge cap with the 5x 10x 40x and 63x objectives By using the software function go to checkpoint the slide is moved out of the light path and the cap can be lowered further towards the objectives for looking inside 7 Downstream Applications 7 1 Preparation of RNA 7 4 1 RNA from frozen sections For capturing microdissected samples from frozen sections we recommend PALM AdhesiveCaps 500 ul opaque 1440 0250 500 ul clear 1440 0255 200 ul opaque 1440 0240 200 ul clear 1440 0245 The RNeasy Micro Kit QIAGEN 74004 in our hands results in a good yield of RNA But in contrast to most other laser microdissection companies you can go on straight forward with RNA extraction by using your extraction procedure of choice After LMPC adjust the sample volume to 350 ul with lysis buffer QIAGEN RLT buffer and incubate upside down for 30 minutes Vortex the digestion mix thoroughly Tissue fragments left undisrupted represent RNA lost Then the lysate is soun down in a bench top centrifuge 5 minutes 13400 rcf e g Eppendorf 5415D 12000 rpm and samples can then be stored f2. PALM AdhesiveCaps Liquid collection other microfuge tubes Capture check looking into the cap to see the catapulted samples Downstream Applications Preparation of RNA RNA from frozen sections RNA from FFPE sections Quality control of RNA General remarks on RNA distribution content RNase activity PALM LabTips for working with RNA Other protocols DNA Chromosomes Live Cells 2 15 PALM MicroLaser Systems Protocols X PALM Microlaser Technologies 1 I ntroduction 1 1 Some remarks on RNA RNA is a biological macromolecule with many different functions Messenger RNA mRNA transcribed from DNA serves as a template for synthesis of proteins This protein synthesis is carried out by ribosomes which consist of ribosomal RNA rRNA and proteins Amino acids for protein synthesis are delivered to the ribosome on transfer RNA tRNA molecules RNAs are also part of riboproteins and ribozymes Analysis of RNA can provide a good reflection of an organism s gene expression profile Gene expression profiling of material isolated by microdissection has become a very important method for analyzing cellular behavior in a micro scale and is used in research and clinical applications Therefore the isolation of high quality RNA is crucial for all subsequent steps and the success of the overall experiment 1 2 The DOs and DON Ts on handling RNA RNA degradation is a common reason for failing experiments RNA is prone
3. done using glass Slides Floating the section on warm water as well as hot plate techniques can be applied After mounting let dry the slides overnight in a drying oven at 56 C To allow cutting and catapulting a coverslip and standard mounting medium must not be applied e Deparaffination Paraffin will reduce the efficiency of the laser sometimes completely inhibiting cutting and catapulting If you are working with unstained sections it is therefore very important to remove the paraffin before laser cutting and pressure catapulting If applying standard staining procedures deparaffination is routinely included in any protocols Minimal procedure 1 Xylene 2 times for 3 10 minutes 2 Ethanol 100 1 minute 3 Ethanol 96 1 minute 4 Ethanol 70 1 minute Note The thin 0 17 mm MembraneSlides are not as resistant against organic solvents and should be handled according to the Minimal procedure see above RNA handling 0406 6 15 PALM MicroLaser Systems Protocols X PALM Microlaser Technologies 3 3 Cytospins Cytospins can be prepared on glass slides or on MembraneSlides After centrifugation with a cytocentrifuge let the cells air dry Then fix for 5 minutes in 100 methanol Allow the cytospins to dry at room temperature before staining 3 4 Blood and tissue smear Distribute a drop of peripheral blood or material of a smear over the slide Be careful to avoid injuries in the membrane which would lead to leakage
4. during fixation or washing steps and therefore will impair the laser pressure catapulting process Let smears air dry shortly and fix them for 2 up to 5 minutes in 70 ethanol 4 Staining procedures For isolation of high quality RNA use only freshly prepared staining solutions and take notice of our tips on handling RNA pp 15 4 1 Frozen sections Most standard histological stainings like HE Methyl Green Cresyl Violet Nuclear Fast Red can be used when you are interested in RNA Note Using frozen sections endogenous RNases may be active after the short fixation step Therefore it is recommended to keep all incubation steps as short as possible Please use RNase free water and solutions for any steps All required reagents should be kept on ice Commonly used protocols are the Cresyl Violet and Hematoxylin Eosin staining 4 1 1 Cresyl Violet This short staining procedure colors the nuclei violet and the cytoplasm weak violet e Procedure After fixation dip slide for 60 seconds in 1 cresyl violet acetate solution Remove excess stain on absorbent surface Dip into 70 Ethanol Dip into 100 Ethanol Air dry shortly 1 2 minutes SP aN Dissolve solid cresyl violet acetate e g ALDRICH 86 098 0 at a concentration of 1 w v in 100 EtOH at room temperature with agitation stirring for several hours to overnight Some unsolubilized powder is normal Filter the staining solution before use RNA handling 0406
5. 64 Thymus 16 Kidney Heart Brain RNA handling 0406 14 15 PALM MicroLaser Systems gt lt z Protocols P A L M Microlaser Technologies 9 PALM LabTips for working with RNA For best RNA quality we use frozen sections on PALM MembraneSlides Frozen sections should not be stored for more than a few days at 80 C Freezing should be performed after staining and drying A prognosis of the expected amount of RNA is difficult to give since many factors will influence the outcome see above From mouse liver frozen sections we usually are able to retrieve 5 20 pg RNA per cell calculated from extractions of 1000 cells and analysis with an Agilent Bioanalyzer Agilent Application Note 5988 EN on our website or at www chem agilent com Note Quantitative results from an analysis with the RNA Pico kit are dependent on the salt content of the sample Archival tissues are often formalin fixed and paraffin embedded RNA extraction from these tissues is not effective because of the cross linking properties of aldehydes Other methodologies for preservation of high molecular weight RNA in FFPE tissue are described by Vincek et al 2005 see website www palm microlaser com publications and Olert et al 2001 Pathol Res Pract 197 823 826 Summarized recommendations e Keep attention to DOs and DON Ts on handling RNA pp 3 e Take PALM AdhesiveCaps as collection device for all RNA experiments pp 9 e Choose a short staining proced
6. 7 15 PALM MicroLaser Systems Protocols X PALM Microlaser Technologies 4 1 2 Hematoxylin Eosin HE HE staining is used routinely in most histological laboratories and does not interfere with RNA preparation The nuclei are stained blue the cytoplasm pink red e Procedure After fixation quickly dip slide 5 6 times in RNase free distilled water Stain 1 minute in Mayer s Hematoxylin solution e g SIGMA MHS 32 Rinse 1 minute in DEPC treated tap water or blueing solution Stain 10 seconds in Eosin Y e g SIGMA HT110 2 32 Perform a quick increasing ethanol series 70 96 100 Air dry shortly OU SVN 4 2 Paraffin embedded FFPE sections After deparaffination continue with the staining procedure of your choice Standard staining procedures can be used for frozen and FFPE sections see above 5 Storage Stained slides can be used immediately or stored at 80 C before LMPC To avoid excess condensation of moisture during thawing the slides should be frozen and rethawed in a tightly sealed container e g 50 ml Falcon tube 6 Laser Microdissection and Pressure Catapulting LMPC Procedures Please additionally have a look at PALM MicroBeam user manual 6 1 Tips for improvement of morphological information For LMPC Laser Microdissection and Pressure Catapulting embedding and glass covering of the specimen is inapplicable Thus the rough open surface of the section material often results in impaired vie
7. RNAs and rRNAs mRNA represents only 1 5 of the total cellular RNA Approximately 360 000 mRNA molecules are present in a single cell corresponding to approximately 12 000 different transcripts with a typical length of 2 kb Some mRNAs comprise as much as 3 of the mRNA pool whereas others account for less then 0 01 QIAGEN Bench guide RNA distribution in a typical mammalian cell Total RNA per cell 10 30 pg rRNA 28S 18S 5S 80 85 tRNAs snRNAs low MW species 15 20 mRNAs 1 5 Total RNA in nucleus 14 DNA RNA in nucleus 2 1 mRNA molecules per cell 2x10 to 1x 10 Typical MRNA size 1900 nt RNA handling 0406 13 15 PALM MicroLaser Systems Protocols X PALM Microlaser Technologies RNA content in various cells and tissues Total RNA ug MRNA yg Cell cultures 107 cells NIH 3T3 120 HeLa 150 COS7 350 Mouse tissue 100 mg Brain 120 Heart 120 Intestine 150 Kidney 350 Liver 400 Lung 130 Spleen 350 NOPRPON DU UIW W Also the RNase activity varies dramatically across different tissues Krosting J Latham G AMBION Inc A comparison of total RNase activities for 8 different mouse tissues showed that total RNase activity spans a 181 000 fold range from pancreas to brain which points out the importance of RNase control Quantitative hierarchy of RNase activity in mouse tissues AMBION Inc Mouse tissues Fold increase relative to brain Pancreas 181 000 Spleen 10 600 Lung 5 300 Liver
8. Stir for 5 6 h at room temperature to dissolve the DEPC 2 Soak the reaction tubes into the DEPC solution take care that the tubes are completely covered with liquid not blistered and incubate overnight at room temperature 3 Autoclave the tubes together with the solution for 20 minutes at 121 C to inactivate the DEPC 4 Discard the liquid carefully and thoroughly Dry the tubes at 50 C 80 C 5 Use the tubes as usual Note DEPC is toxic and should be used under a hood 6 3 Collection procedures Please have a look at PALM MicroBeam user manual 6 3 1 Dry collection PALM AdhesiveCaps Note P A L M recommends PALM AdhesiveCaps as collection device for all RNA experiments Capturing without liquid minimizes RNase activity After LMPC add lysis buffer of your own choice e g QIAGEN 350 ul RLT buffer and centrifuge the lysis mix after an upside down incubation 30 minutes AWW Then the routine RNA extraction procedure can be attached RNA handling 0406 10 15 PALM MicroLaser Systems Protocols X PALM Microlaser Technologies 6 3 2 Liquid collection other microfuge tubes Pipette 20 ul lysis buffer into the cap The catapulted cells or cell areas will stick onto the wet inner surface of the cap and will not fall down after the catapulting procedure Be aware that aqueous solutions will dry out after a while When using glass mounted samples it may be advisory to put more liquid up to 40 ul
9. X PALM Microlaser Technologies 2 25 D S amp S 40 4 5 5 6 amp s wild RNA Area 125 8 RNA Concentration 695 pgp rRNA Ratio 28x 18x 19 RNA Integrity Number RIN 10 Laser Microdissection and Pressure Catapulting LMPC PROTOCOLS RNA Handling Contents PREP PPP PB N NNPRPRPH WNE AUNE m QUARARRWWWWWNNNNNNNNE EE NBER N e ADAHAD NNER m WNE N AWWW N e N e NNNNYADAAG Nere 10 1 10 2 10 3 RNA handling 0406 A P A L M Microlaser Technologies Introduction Some introducing remarks on RNA The DOs and DON Ts on handling RNA First steps of sample preparation Preparation of slides Samples on PALM MembraneSlides Treatment to remove RNases UV treatment Poly L Lysine treatment Samples on glass slides Treatment to remove RNases Mounting samples onto slides Frozen sections Paraffin embedded FFPE sections Cytospins Blood and tissue smear Staining procedures Frozen sections Cresyl Violet Hematoxylin Eosin HE Paraffin embedded FFPE sections Storage Laser Microdissection and Pressure Catapulting LMPC Procedures Tips for improvement of the morphological information PALM Diffusor CM 1210 0320 PALM AdhesiveCaps 1440 0240 1440 0250 PALM LiquidCover Glass N 1440 0600 Collection devices PALM AdhesiveCaps 1440 0240 1440 0250 1440 0245 1440 0255 Other microfuge tubes Collection procedures Dry collection
10. ed RNase activity For more details and handling please see PALM LiquidCover Glass N instruction manual 6 2 Collection devices 6 2 1 PALM AdhesiveCaps 500 ul opaque 1440 0250 500 ul clear 1440 0255 200 ul opaque 1440 0240 200 ul clear 1440 0245 The intention of PALM AdhesiveCaps is to allow LMPC Laser Microdissection and Pressure Catapulting without applying any capturing liquid into the caps prior to LMPC This minimizes RNase activity Beside the quick relocation of the catapulted samples in the cap due to instant immobilization there is no danger of evaporation and crystal forming during extended specimen harvesting Furthermore the filling of the caps allows a low energy catapulting due to the short distance between slide and collection device For more details and handling please see also PALM AdhesiveCaps product information Note P A L M recommends AdhesiveCaps as collection device for all RNA experiments RNA handling 0406 9 15 PALM MicroLaser Systems Protocols X PALM Microlaser Technologies 6 2 2 Other microfuge tubes In case you don t like to work with PALM tubes use commercially available RNase free plasticware e g ABgene AB 0350 0 5 ml tubes If there are only no RNase free tubes available use the following procedure to remove RNases e Treatment of microfuge tubes to remove RNases Add 0 1 ml DEPC to 100 ml of double distilled water to get a 0 1 DEPC solution DEPC e g ROTH K028 1 1
11. lution to the 20 ul of the digested lysate from above vortex thoroughly and transfer to a 1 5 ml microfuge tube 2 Incubate at room temperature for 30 minutes 3 Add 100 ul of protein precipitation solution mix carefully by inverting the tube at least ten times 4 Incubate on ice for 30 minutes 5 Centrifuge at maximum speed for 15 minutes in a cooled centrifuge e g Eppendorf 5417C 14000 rpm 4 C 6 Carefully transfer the supernatant into a fresh tube Do not disturb the white DNA protein pellet Discard the pellet together with the used tube 7 Add 0 5 ul of a glycogen solution 20 mg ml as carrier substance to the transferred supernatant 8 Add 300 ul isopropanol and mix carefully but thoroughly by inverting the tube fifty times 9 Centrifuge for 25 minutes as above 4 C max speed Mark the orientation of the tubes in the rotor 10 Remove supernatant carefully and discard it The pellet is mostly not visible Look for the mark on the tube and remove the liquid with a thin pipette tip on the opposite side of the expected pellet position RNA handling 0406 12 15 PALM MicroLaser Systems Protocols X PALM Microlaser Technologies 11 The pellet is washed briefly by adding 200 ul of pre cooled ethanol 70 20 C 12 Centrifuge for 15 minutes as above 4 C max speed Keep the orientation of the tubes in the rotor as before 13 Remove all supernatant carefully and discard it The pellet should
12. now be visible as a small point of about 1 mm remove the liquid with a thin pipette tip on the opposite side of the pellet position 14 The pellet is now air dried for 20 30 minutes at room temperature Do not use vacuum drying and then resolved in 10 ul of RNA Hydration Buffer for at least 30 minutes at 4 C or in ice The RNA solution can now be used for cDNA synthesis or stored at 20 80 C We normally use half of the solution in a RT reaction of 20 ul e g Transcriptor First Strand cDNA Synthesis Kit ROCHE 04 379 012 001 using random oligomers instead of oligoT as primers for the cDNA synthesis 7 2 Quality control of RNA The most common method used for assessing the integrity of total RNA is to run the RNA sample on an agarose gel In general at least 200 ng of RNA must be loaded onto the gel To analyze RNA samples with concentrations down to 50 pg ul the Agilent 2100 Bioanalyzer is an alternative to traditional gel based analysis and provides information about RNA quality degradation purity and quantity www chem agilent com A prognosis of the expected amount of RNA in a tissue is difficult to give since many factors like species cell tissue type fixation staining fragmentation extraction procedure and others will influence the outcome 8 General remarks on RNA distribution content RNase activity A typical mammalian cell contains 10 30 pg total RNA mRNA rRNA tRNA The majority of RNA molecules are t
13. o overcome the hydrophobic nature of the membrane it is advisable to irradiate with UV light at 254 nm for 30 minutes e g in a cell culture hood The membrane gets more hydrophilic therefore the sections paraffin and cryosections adhere better Positive side effects are sterilization and destruction of potentially contaminating nucleic acids 2 1 1 3 Poly L Lysine treatment Additional coating of the slide with Poly L Lysine 0 1 w v only will be necessary for poorly adhering materials e g brain sections and should be performed after UV treatment Distribute a drop of the solution on top of the membrane Let air dry at room temperature for 2 3 minutes Avoid any leakage underneath the membrane as this might result in impairment of Laser Pressure Catapulting 2 1 2 Samples on glass slides With PALM MicroBeam almost every kind of biological material can be microdissected and catapulted directly from glass slides Even archival pathological sections can be used after removing the cover slip and the mounting medium To facilitate easy catapulting additional adhesive substances or Superfrost charged slides should only be applied when absolutely necessary for the attachment of poorly adhering special material e g some brain sections or blood vessel rings For catapulting higher laser energy is needed 2 1 2 1 Treatment to remove RNases Treatments of glass slides to remove RNases are identic with the procedure for PALM Memb
14. or later use at 80 C RNA handling 0406 11 15 PALM MicroLaser Systems gt lt Protocols P A L M Microlaser Technologies 7 1 2 RNA from FFPE sections For capturing of microdissected samples we recommend PALM AdhesiveCaps 500 ul opaque 1440 0250 500 ul clear 1440 0255 200 ul opaque 1440 0240 200 ul clear 1440 0245 Note For FFPE samples an additional Proteinase K digestion step is necessary Do not forget to inactivate the Proteinase K after the digestion 90 C for 10 minutes After LMPC add 20 ul of lysis buffer containing Proteinase K 150 mM NaCl 100 mM Tris pH 7 5 0 5 Igepal 0 5 ug ul Proteinase K directly onto the microdissected sample which is still located in the cap Proteinase K digestion is performed in an upside down position pp 10 at 55 C overnight in an incubator to avoid evaporation of the lysate The time necessary for complete digestion depends on the kind and on the number of catapulted cells If not proceeding immediately store the samples at 20 C or 80 C All components of the following extraction procedure except glycogen are from the Purescript RNA isolation kit GENTRA USA 212010 Mainly follow the standard protocol of the Purescript Kit This procedure is very simple and allows high end concentrations of RNA due to an effective precipitation step Also some loss of the minute RNA amounts on columns is avoided Protocol in brief 1 Add 300 ul cell lysis so
15. pt the working distance to the different glass slides by moving the correction ring on the objective Due to the short working distance of the 100x magnifying objectives only 0 17 mm thin cover glass slides can be used Note The thin 0 17 mm MembraneSlides are not resistant against heat Use other treatments to remove RNases PALM MembraneSlides NF nuclease free are certified to be free of DNase RNase and human DNA In addition to PEN MembraneSlides P A L M offers for special processes i e for isolation of chromosomes PET or POL membrane covered slides If you need information about these slides please contact salesteam palm microlaser com RNA handling 0406 4 15 PALM MicroLaser Systems Protocols X PALM Microlaser Technologies 2 1 1 1 Treatment to remove RNases PALM MembraneSlides are shipped without any pretreatment e To ensure RNase free MembraneSlides heat MembraneSlides at 180 C for 4 hours to completely inactivate RNases e An alternative for decontamination of slides is the treatment with RNase ZAP AMBION 9780 First dip the slides for a few seconds into pure RNase ZAP followed by two separate washings in DiEthyIPyroCarbonate DEPC treated distilled water and drying at 37 C for 30 minutes up to 2 hours e PALM MembraneSlides NF nuclease free are certified to be free of DNase RNase and human DNA Using these slides treatments to remove nucleases are not necessary 2 1 1 2 UV treatment T
16. raneSlides 2 1 1 1 RNA handling 0406 5 15 PALM MicroLaser Systems Protocols PALM Microlaser Technologies 3 Mounting samples onto slides 3 1 Frozen sections e Sectioning Sections are mounted onto MembraneSlides the same way as routinely done using glass slides To allow subsequent cutting and catapulting a coverslip and standard mounting medium must not be applied Freezing media like OCT or similar may be used but should be kept to a minimum and have to be removed before laser cutting For optimal RNA retrieval take a pre cooled MembraneSlide warm the back of the slide shortly with your finger gloves transfer section as routinely done by touching with the warmed area and let the slide remain in the cryostat for 2 3 minutes e Fixation After mounting the sections there are many possibilities to fix the material P A L M recommends the dehydration in ice cold 70 ethanol for 2 3 minutes e Removing the tissue freezing medium If OCT or another tissue freezing medium is used it is important to remove it before Laser Microdissection because these media will interfere with laser efficiency Removing of the medium is easily done by dipping the slide 5 6 times in ice cold RNase free water If the sections will be stained the supporting substance is normally removed automatically in aqueous staining solutions 3 2 Paraffin embedded FFPE sections Sections are mounted onto MembraneSlides the same way as routinely
17. to digestion by a wide variety of endogenous and exogenous RNases These RNases are present on almost all objects that come into contact with human skin and are difficult to inactivate Even minute amounts are sufficient to destroy RNA Some precautions can make the difference between an intact and degraded RNA prep www ambion com and therefore between successful and unsuccessful experiments DOs e Designate a special area for working with RNA e Clean benches with special cleaning solutions e g RNaseZap AMBION 9780 e Wear gloves and change them frequently e Use sterile disposable plasticware e Glassware should be treated with DiEthyIPyroCarbonate DEPC and oven baked at 180 C for at least 4 hours before use e Use filtered pipette tips e Solutions water or other solutions should be prepared with 0 1 DEPC e Use RNase free reagents tubes and tips e For best results use either fresh samples or samples that have been snap frozen on dry ice or in liquid nitrogen all required reagents should be kept on ice e Store prepared RNA aliquoted in ethanol or RNA elution buffer at 80 C e To avoid condensation of moisture during thawing the slides should be frozen at 80 C and rethawed in a tightly sealed container e g 50 ml Falcon tube e In general use protocols e g staining with short incubation times on ice DON Ts e Don t breath on samples some researchers wear masks e Don t touch anything with bare hands e Don t a
18. ure for tissues with high content of endogenous RNases e g Cresyl Violet pp 7 e QIAGEN RNA extraction kit results in good RNA yield quality and quantity from frozen sections in our lab pp 11 e GENTRA RNA extraction kit results in good RNA yield quality and quantity from FFPE tissue in our lab pp 12 10 Other protocols 10 1 DNA DNA protocols on request 10 2 Chromosomes Protocols on request 10 3 Live cells Live cell protocols on request Please note There is also a review brochure available dealing with laser micro manipulation of live cells P A L M Scientific Edition No 11 ISBN No 3 9808893 0 0 For questions comments or protocol requests please contact PALM Application Laboratory Email lab palm microlaser com Service Line 49 0 81 58 99 71 300 RNA handling 0406 15 15
19. utoclave pipette tips as water vapor may contain RNases e Don t allow frozen tissue to thaw e Don t resuspend RNA in DEPC water residual DEPC can inhibit downstream reactions RNA handling 0406 3 15 PALM MicroLaser Systems Protocols X PALM Microlaser Technologies 2 First steps of sample preparation 2 1 Preparation of slides 2 1 1 Samples on PALM MembraneSlides PEN membrane covered 1 mm 1440 1000 PEN membrane covered 0 17 mm 1440 1500 PEN membrane covered 1 mm nuclease free 1440 1600 MembraneSlides are special slides covered with a membrane on one side This membrane is easily cut together with the sample and acts as a stabilizing backbone during catapulting Therefore even large areas are catapulted by a single laser impulse without affecting the morphological integrity Use of MembraneSlides is especially important for isolating single cells chromosomes as well as living cells or small organisms P A L M offers slides 1 mm 0 17 mm covered with polyethylene naphthalate PEN membrane This PEN membrane is highly absorptive in the UV A range which facilitates laser cutting The membrane can be used for all kind of applications When working with low magnifying objectives like 5x 10x or 20x regular 1 mm thick glass slides and 0 17 mm glass slides can be used To keep this flexibility for higher magnifications 40x or 63x P A L M recommends using long distance objectives With those you have the possibility to ada
20. w of morphology 6 1 1 PALM Diffusor CM 1210 0320 PALM Diffusor CM can be inserted into PALM CapMover as any holder and swivels over the sample The opaque glass diffuses the incident microscope light which smoothens the harshness of contrast and depending on material and staining even minute details as nuclei and cell boundaries show up Even slight differences in color become visible For more details and handling please see PALM Diffusor CM instruction manual RNA handling 0406 8 15 PALM MicroLaser Systems Protocols X PALM Microlaser Technologies 6 1 2 PALM AdhesiveCaps 500 ul opaque 1440 0250 200 ul opaque 1440 0240 The white opaque filling of PALM AdhesiveCaps clearly improves visualization of morphological information of the samples at the object plane due to enhanced color balance and contrast which makes the view comparable to those of coverslipped tissue sections Two different microfuge tube sizes with these filled caps are available from P A L M For more details and handling please see PALM AdhesiveCaps product information 6 1 3 PALM LiquidCover Glass N 1440 0600 The polymeric and low viscose PALM LiquidCover Glass N completely embeds the tissue and smoothens the rough tissue surface resulting in enhanced morphology The coverslip like surface that is formed not only improves the optical characteristics of the specimen but also protects it against environmental influences e g moisture and associat
Download Pdf Manuals
Related Search