CytoViva User Manual
Contents
1. 1 cover glass 1mm standard microscope slides i LabTek Chambers slides have been used successfully i Total Focal depth of CytoViva is 1 5mm Mono layer sub confluent cells are ideal samples If you are using tissue samples the ideal thickness is 30 microns or less All sample preparation must be evaluated on an individual case Limit debris or air bubbles in the field of view Use clean cover glass and slides Depending on specific applications sample volumes of 4 10 ul have been Shown to work best 2 Oil Immersion for best results use MaA a Type A immersion oil nd gt 1 515 b Ensure there is enough oil to completely cover the entire illuminated surface of the CytoViva lens when in contact with underside of sample Do not mix oil types on samples Do not allow water or other material fluids to mix with oil User may need to add some oil between samples After viewing sample dab oil off of objective with lens cloth no need to remove oil from CytoViva between samples Oils specific for fluorescence microscopy may improve fluorescent observation 3 Objectives a b Q h g A variety of objectives will work with CytoViva Results may vary depending on the installed objective set Higher quailty objectives will likely produce better images The NA of the objective must be below 1 2 NA of objective can adjust brightness reduce glare or improve contrast i Note decreasing NA of the objective w2. Manual Warranty Card O O O O O OO O Part 3 Parts of the oViva Adapter 1 CytoViva Adapter Periscope Head raised Dovetail Groove and Ring Filter Slot Light Guide Port PANE 1 CytoViva Adapter End 2 Light Source End 3 Light Source 1 Light Guide Port 2 Light Source Diaphragm a Left less light b Right more light Part 4 CytoViva Installation 1 CytoViva Installed a Periscope raised operational position Ve ify H iU it a Beastie i P a i betas iF r b Periscope lowered for installation purposes 2 CytoViva Installation CytoViva installs on most research grade microscopes Adapter kits have been made for specific models and are designed on an individual basis a Determine the type of condenser mount on your microscope see below o C shaped example o O shaped example o 3 point example b Then follow the appropriate installation instructions The possible types of condenser mounts include o C Shaped Condenser Mount common on Olympus o O Shaped Condenser Mount common on Nikon o 3 Point Condenser Mount common on Zeiss 1 C Sha 1 2 3 4 5 6 1 8 Condenser Mount common on Olympus Raise specimen stage using course focus Using the condenser focus know position the condenser mount 2 3 from top of travel Remove installed condenser Lower periscope on CytoViva Insert CytoViva dovetail into C mount a Shou
3. A coarse adjustment knob filter selection 3 A fine adjustment knob 4 Light guide to CytoViva adapter 12 2 Liquid Light Guide Between CytoViva Adapter and DMF Module 1 CytoViva Adapter End 2 DMF Module End Front Port 3 Liquid Light Guide Between Solarc Light Source and DMF Module 4 Liquid Light Guide Between EXFO light Source and DMF Module 13 5 Light Sources a Solarc 24W Metal Halide Light Source i Included with the CytoViva Adapter 1 Light guide port Note no diaphragm when used with DMF module to improve intensity 2 Power button b EXFO X CITE 120 Q Light Source Optional i Recommended when using Fluorescence 1 Power button 2 Iris Shutter dial Note Consult Exfo manual for operation When using the Exfo light source in combination with the CytoViva DMF a UV blocking emission filter must be used 14 Part 9 DMF Installation 1 Light Source a The system is compatible with a Solarc light source and the EXFO X Cite 120 b CytoViva DMF is standard equipped with a Solarc unit c A Solarc light source can be used for basic fluorescence techniques however considering the intensity and spectral characteristics required for most fluorescence it is recommended that the EXFO Xcite 120 be used NOTE When using the Exfo X Cite 120 in combination with the CytoViva DMF a UV blocking emission filter must be used 2 Light Guide a To set up the system take the light guide fr
4. ANTY Additional Notes on Use 1 Do not look directly into light source adapter or light guide 2 Do not open light source case while light source is connected to power supply 3 Protect light source from liquids 4 Protect the CytoViva lens surface As with any microscopic lens scratches will diminish performance 5 Protect CytoViva as you would any other microscopic component It is designed for normal microscopic use 6 Do not make modifications or adjustments to the CytoViva adapter not outlined in this document 7 Do not kink or sharply bend the liquid light guide 8 Use of the Exfo light source in combination with the CytoViva system requires the use of a UV blocking emission filter 11 Part 6 CytoViva Dual Mode Fluorescence DMF Module Overview The CytoViva Dual Mode Fluorescence DMF module enables researchers to easily modulate simultaneously between multiple fluorescent and non fluorescent labels or any combination of the modes Samples are viewed directly through the microscope eyepiece in real time and quickly captured by a standard microscope camera in a single exposure Part 7 The Standard CytoViva DMF system includes o CytoViva DMF Module o Light Source Light Guide o Light Source zs Solarc Light Source Optional Exfo Light Source Part 8 Parts of the CytoViva DMF Module 1 CytoViva Dual Mode Fluorescence DMF Module pian ae 1 Four filter locations numbered 1 4 indicating mode 2
5. IMPORTANT PLEASE READ CAREFULLY t Viva Limited Warranty CytoViva warrants for a period of one 1 year from the date of purchase from CytoViva Inc or an authorized agent of CytoViva Inc the Warranty Period that the unmodified CytoViva adapter and or Dual Mode Fluorescence Module the Equipment when new and subject to normal use and service shall be free of defects in materials and workmanship and shall perform in accordance with the manufacturer s specifications If any component of the Equipment does not function properly during the Warranty Period due to defects in material or workmanship CytoViva will at its option either repair or replace the component without charge subject to the conditions and limitations stated herein Such repair service will include all labor as well as any necessary adjustments and or replacement parts If replacement components are used in making repairs these components may be remanufactured or may contain remanufactured materials Repair or replacement without charge is CytoViva s only obligation under this warranty This warranty is NOT transferable from the original purchaser of the Equipment Limitations Other components of the product package specifically including but are not limited to the light source s light source power transformer s and cord s liquid light guide s optical filters spectrophotometer camera s software microscope part s and motorized stage are w
6. Viva it should be returned postage paid along with the original dated receipt to CytoViva Inc 300 North Dean Road Suite 5 PMB 157 Auburn AL 36830 Your repaired item or replacement product will be returned to you postage paid In the event the purchaser returns Equipment to CytoViva and it is determined by CytoViva that the Equipment has been returned without cause the purchaser will be notified and the Equipment returned at the purchaser s expense DI SCLAI MER OF WARRANTIES LIMITATION OF LIABILITY THE WARRANTIES CONTAINED HEREIN ARE IN LIEU OF AND CYTOVIVA EXPRESSLY DISCLAIMS AND CUSTOMER WAIVES ALL OTHER REPRESENTATIONS AND WARRANTIES EXPRESS OR IMPLIED STATUTORY ARISING IN THE COURSE OF DEALING OR PERFORMANCE CUSTOM USAGE IN TRADE OR OTHERWISE INCLUDING WITHOUT LIMITATION IMPLIED WARRANTIES OF MERCHANTABILITY TITLE INFRINGEMENT OR FITNESS FOR A PARTICULAR PURPOSE WHERE NOT PROHIBITED BY LAW CYTOVIVA WILL NOT BE RESPONSIBLE FOR ANY CONSEQUENTIAL OR INCIDENTAL DAMAGES RESULTING FROM THE PURCHASE USE OR IMPROPER FUNCTIONING OF THIS EQUIPMENT REGARDLESS OF THE CAUSE SUCH DAMAGES FOR WHICH CYTOVIVA WILL NOT BE RESPONSIBLE INCLUDE BUT ARE NOT LIMITED TO LOSS OF REVENUE OR PROFIT DOWNTIME COSTS LOSS OF USE OF YOUR EQUIPMENT COST OF ANY SUBSTITUTE EQUIPMENT FACILITIES OR SERVICES OR CLAIMS OF THIRD PARTIES FOR SUCH DAMAGES CYTOVIVAS LIABILITY WILL NEVER EXCEED THE PURCHASE PRICE OF ANY DEFECTIVE PRODUCT OR PART Thank y
7. adjust condenser focus knob d Insert polarizing filter e If available adjust objective iris Fluorescence a No signal or weak signal b Sample may be bleached c Fluorescence label may not be present d Excitation or emission filters may not correspond to fluorochromes For support issues please call or email support cytoviva com 1 888 737 3130 19
8. arranted based on the individual original manufacturer s warranties and policies There is no warranty whatsoever on the contrast filters bulbs or the coil on the motorized stage purchased as part of the Equipment This warranty does not cover circumstances beyond CytoViva s control breakage or a malfunction that has resulted from improper or unreasonable use or maintenance accident tampering misuse neglect improper installation modification improper maintenance or service cleaning procedures shipping or repacking of Equipment or service or parts to correct problems where such service or parts are performed or provided by anyone other than CytoViva or an authorized agent of CytoViva service required as the result 6 unauthorized modifications or service misuse or abuse failure to follow CytoViva s operating maintenance or failure to use items supplied by CytoViva This warranty is also void if the light source light source power transformer and cord or liquid light guide is not used in accordance with the original manufacturer s instructions recommendations or documentation Warranty service will not be provided without a dated proof of purchase Please return the Warranty Registration Card together with a copy of the original receipt within thirty 30 days of purchase It is the purchaser s responsibility to return the Equipment to the authorized agent from whom it was purchased If the Equipment was purchased directly from Cyto
9. e light path this arrow should point toward the front of the unit Gently snug the set screw and replace the housing 17 H a ee CytoViva System CytoViva Adapter CytoViva DMF Module Liquid Light Guides Light Source optional Exfo pictured Emission Filter Tray Note Emission filter must be in filter tray 18 Part 13 Troubleshooting l Light Source Will Not Turn On a The light source is equipped with a thermal auto off switch to protect overheating Allow the light source to cool down with power off for 15 minutes and try to power on b Check that power supply LED light is illuminated and the power supply is plugged into the light source c The light bulb may be burned out and should be replaced No Light on the Sample a Pull the Periscope to the raised position on the CytoViva Adapter The I mage is Blurry or Distorted a Ensure CytoViva is properly aligned and focused b Sample may be too thick or granular Oil may have been contaminated with water Clean oil off of CytoViva lens objective and sample Reapply oil d Sample glass may be dirty Clean glass surfaces 2 There is Debris in the Field a Reduce volume of sample buffer b If possible wash your sample with clean media or buffer prior to mounting c Sample glass may be dirty Clean glass surfaces The I mage is too Bright a Reduce light source diaphragm b There may be too much debris in the field c Slightly
10. each filter b The fine adjustment knob can be used to transition between filters 4 The key feature of the system is that the user can easily transition between any combination of full fluorescence and unfiltered modes 15 Part 11 DMF Mode Selection Using the fine adjustment knob to transition between modes Note Transitioning to the shutter mode from the full fluorescent or full unfiltered modes will dim the respective mode Modes Dial Position Full Fluorescent Mode Partial Unfiltered Mode L Partial Fluorescent Mode a j Full Unfiltered Mode Shutter Mode 16 Part 12 Filter Installation and Ordering I nformation Consult with your CytoViva representative to help select the appropriate filter sets The DMF module uses 25mm diameter exciter filters 1 To install filters q b Remove the painted housing of DMF module by removing the 8 screws on the sides of the unit The filter position indicated by the dial corresponds to the filter holder located 180 on the wheel e g 12 o clock position in the dial corresponds to the 6 o clock position on the wheel Filters are held in place by a single screw on the edge of the wheel i Note Filter corresponding with dial position is inline with optic path Loosen the desired location set screw and remove existing filter i Note Use a piece of lens paper to remove and install filters to avoid damaging the filter surfaces Install new filter noting th
11. ill decrease resolution adjust as your application merits Not all objectives have an iris Most 60x 100x objectives will require an adjustable iris NA Objectives with a NA of gt 1 2 require an iris Long distance 40x objectives have been used with good results 4 Recommended Objectives a b C Plan Fluorite 10x air Plan Fluorite 40x 0 75 NA or 40X oil with iris 60x 100x oil with tris 1 35 1 00 NA 5 Light Source Diaphragm Only on units without fluorescence module small lever on the light guide mount of the light source rotate left and right q Can be adjusted to increase or decrease illumination 10 6 Adapter Filters a Three included filters red green and polarizing filter b Polarizing filter may improve the image quality of certain samples c Use as you feel appropriate these filters have no fluorescent properties 7 Using Condenser Focus a A slight focus adjustment of CytoViva may be used to regulate image brightness or glare i Note use care not to push CytoViva into sample slide 8 Cleaning and Care a Clean the CytoViva lens and objective after use every time i Use high quality lens cleaner and lens paper to prevent residue buildup b Wipe any excess oil from the CytoViva body c No mechanical adjustment or maintenance of the CytoViva adapter is required or recommended DO NOT REMOVE OR ADJ UST FASTENERS ON THE CYTOVIVA ADAPTER DOI NG SO MAY DAMAGE THE ADAPTER AND WILL VOID THE WARR
12. ld be engaged a full 180 of CytoViva dovetail Light guide port can be in any convenient orientation Lightly tighten set screw Do not over tighten Raise periscope on CytoViva Make no contact with stage ii O Shaped Condenser Mount common on Nikon 1 2 3 Ot 1 8 9 Raise specimen stage using course focus Remove installed condenser Raise condenser mount fully a In some cases the condenser mount has a small silver stop on the top left of the condenser mount that limits the condenser travel This stop is a spring loaded plunger that is locked in place with a set screw Loosen the set screw to allow full travel of the condenser Lower periscope on CytoViva Remove CytoViva dovetail adjustment ring unscrews Position CytoViva into condenser mount so threads extend just above mount Replace CytoViva dovetail adjustment ring Lightly tighten so that CytoViva does not wobble in mount Raise periscope on CytoViva taking care not to contact underside of stage iii 3 Point Condenser Mount common on Zeiss 1 2 3 4 or 8 9 10 Raise stage using course focus Remove installed condenser Remove condenser mount from microscope stand Remove CytoViva dovetail adjustment ring not needed for this application Fully loosen the alignment screws Align rear spring loaded mounting pin of condenser mount with the small pin groove of CytoViva Groove is visible once the dovetail adjustment
13. om the light source and fully insert it into the rear port of the CytoViva DMF unit and tighten thumb screw b Fully insert the light guide from the CytoViva adapter to the front port of the DMF unit gently tightening the thumb screw c Turn on the light source i Do not kink or fold the light guide a gentle loop is acceptable d Attach the light source to the power supply The light source may require a few minutes to warm up and Stabilize Slight color fluctuations may be observed 3 Emitter Filter CONSULT WITH CYTOVIVA REPRESENTATIVE FOR DETAILS a The position of the filter may vary depending on the microscope b The filter may be in a filter tray or a filter cube c When using the Exfo in combination with the CytoViva system a UV blocking emission filter must be used Part 10 DMF Use and Adjustments 1 CytoViva Adapter Focus Alignment a Alignment for this system is described in the CytoViva Adapter Installation Part2 3 b Alignment should be conducted with the DMF module in the full unfiltered mode See below DMF Mode Selection 2 Coarse Adjustment Knob a The coarse adjustment knob allows for convenient selection of filters b There are four filter locations on the module indicated by the numbered positions These numbered positions correspond to the full fluorescence mode of the specific filter 3 Fine Adjustment Knob a The fine adjustment knob is used for precisely modulating between modes of
14. ou for purchasing CytoViva products Table of Contents Chapter 1 CytoViva amp Dual Mode Fluorescence Module User Manual Part 1 CytoViva Adapter Overview Part 2 Standard CytoViva System Components Part 3 CytoViva Adapter Components Part 4 CytoViva Installation Part 5 CytoViva Use and Adjustment Part 6 CytoViva Dual Mode Fluorescence DMF Module Overview Part 7 The Standard CytoViva DMF system includes Part 8 CytoViva DMF Module Components Part 9 DMF Installation Part 10 Using and Adjusting the DMF System Part 11 DMF Mode Selection Part 12 Filter Installation and Ordering Information Part 13 Troubleshooting CHAPTER 1 CytoViva Optical Microscope User Manual Part 1 CytoViva Adapter Overview CytoViva is an advanced illumination system that attaches to research grade microscopes It mounts in place of the standard microscope condenser CytoViva when properly aligned provides a high contrast image with high resolution and a low limit of detection This manual is designed to allow a moderately experienced microscopist to install align use and adjust the CytoViva system Description for installation and use of the optional CytoViva Dual Mode Fluorescence DMF system begins on page 12 Part 2 The Standard CytoViva system includes CytoViva Adapter Liquid Light Guide Light Source Adapter Lens Protector optional during use Filters polarizer red green CytoViva Installation Manual this document Light Source
15. ow 1 If not your sample will not receive illumination Attention After installation manually raise periscope to upright position l Periscope in stop position 3 Alignment i Turn on the light source power 1 Note If you are using the optional Exfo light source an emission filter capable of blocking ultraviolet UV light must be used i Apply 3 large drops of Type A immersion oil to the CytoViva lens Avoid air bubbles in the oil Air bubbles can be dragged off illuminated lens surface with the tip of the oil applicator Air bubbles will create undesirable noise in the image iii Place sample slide on stage iv Raise CytoViva using the condenser focus knob so the oil contacts lower surface of the slide v Using a low power objective e g 4x or 10x focus the objective on the sample see Figure 1 vi Use the condenser focus knob to focus CytoViva on the sample See Figure 2 CytoViva is focused when the center spot of illumination is the smallest vil Use the condenser alignment screws to center the CytoViva illumination spot see Figure 3 1 Note As with conventional condensers slight realignment may be required after changing objectives Figure 1 CytoViva Illumination Figure 1 CytoViva Illumination Not Focused Focused Not Aligned Not Aligned Figure 1 CytoViva Illumination Focused Aligned 9 Part 5 CytoViva Use and Adjustment 1 Samples for best results use a b No
16. ring is removed Gently compress spring loaded pin with CytoViva and align pin groove with alignment screws The alignment screws should mate with the pin groove Alignment screws may need to be adjusted to ensure proper mating Adjust alignment screws so CytoViva is approximately centered Replace condenser mount onto microscope stand taking care to protect CytoViva lens surface Raise periscope on CytoViva c Attach Liquid Light guide to the CytoViva Adapter i Insert black metal adapter of light guide into CytoViva light guide port aligning V groove on adapter with the thumb screw on the port i Insertion stop on adapter should seat tightly against port loosen port thumb screw f required iii Lightly tighten port thumb screw should align with V Groove of adapter Adapter should now be stable no rotation in out movement or wobble d Attach Liquid Light Guide to Light Source i Insert silver adapter of light guide into light source port until adapter stop is seated against port face i No prerequisite orientation required iii Lightly tighten light source port thumb screw 1 Do not kink or fold light guide a gentle loop of guide is acceptable 2 Attach light source to the 12V power supply 3 Light source may require a few minutes to warm up and Stabilize Slight color fluctuations may be observed e CytoViva Periscope i Double check to make sure that the CytoViva periscope is in the raised position see bel
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