BDD-224 International CytoRich non-gyn User Manual
Contents
1. Red as a fixative Was h IN Qs for Bladder washings see Urine protocol gt Submit fresh unfixed specimens to the laboratory immediately after collection or gt Fix specimen with an equal volume of BD CytoRich Red preservative when transport to the laboratory or processing is delayed BD CytoRich non gyn processing procedure gt If fresh unfixed Follow General Procedure Steps I IV Concentrate Fix Wash Process gt If received fixed Follow General Procedure Steps IV If sample is received fixed in BD CytoRich Red skip Step Il Brushings gt Make direct smear s if desired then immediately place brush into approximately 10 mL of BD CytoRich Red preservative or gt Immediately place brush into approximately 10 mL of BD CytoRich Red preservative BD CytoRich non gyn processing procedure gt Remove brush after vortexing but before centrifugation steps gt If received fresh unfixed Follow General Procedure Steps IV Concentrate Fix Wash Process gt If received fixed Follow General Procedure Steps IV If sample is received fixed in BD CytoRich Red skip Step Il Body Cavity Fluids gt Large volume specimens 100 mL Submit fresh specimen to laboratory immediately after collection If transport to laboratory or processing is delayed refrigeration is recommended gt Small volume specimens lt 100 mL Submit fresh specimen to the laboratory immediately2. training For additional questions after installation contact Technical Support BD Diagnostics TriPath No part of this publication may be reproduced transmitted transcribed stored in a retrieval system or translated into any language or computer language in any form or any means electronic mechanical magnetic optical chemical manual or otherwise without the prior written permission of BD Diagnostics TriPath 780 Plantation Drive Burlington North Carolina 27215 United States of America Although this document has been prepared with precautions to ensure accuracy BD Diagnostics TriPath assumes no liability for any errors or omissions nor for any damages resulting from the application or use of this information Information presented in this manual refers to specific applications and protocols for use of non gynecological samples on the BD PrepStain system Specific instructions regarding preservative fluids may be found in the BD CytoRich preservative package inserts BD BD Logo and all other trademarks are property of Becton Dickinson and Company 2007 BD 980 06789 01 Rev A 10 07 For Use Outside the US Only BD BD Diagnostics TriPath Ikaros Business Park lkaroslaan 73 1930 Zaventem BELGIUM Phone 32 3 704 43 80 Fax 32 2 721 36 00 Email TriPathEurope BD com www tripathimaging com www BD com
3. after collection If transport to laboratory or processing will be delayed fix specimen with an equal volume of BD CytoRich Red preservative BD CytoRich non gyn processing procedure gt If fresh unfixed Follow General Procedure Steps I IV Concentrate Fix Wash Process gt If received fixed Follow General Procedure Steps IV If sample is received fixed in BD CytoRich Red skip Step Il Sputum gt If received fresh promptly fix in equal volumes of BD CytoRich Blue preservative gt If transportation to laboratory is delayed collect directly into 30 mL BD CytoRich Blue preservative and send to lab BD CytoRich non gyn processing procedure gt If fresh unfixed Follow General Procedure Steps I IV Concentrate Fix Wash Process gt If received fixed Follow General Procedure Steps I IV If sample is received fixed in BD CytoRich Blue skip Step Il Urine Bladder Washing gt If received fresh fix specimen with an equal volume of BD CytoRich Blue preservative gt Collect specimen directly into 30 mL of BD CytoRich Blue preservative BD CytoRich non gyn processing procedure gt Follow General Procedure Steps IV If sample is received fixed in BD CytoRich Blue skip Step Il Fine Needle Aspirates gt Make direct smear s if desired then immediately rinse the needle in approximately 10 mL of BD CytoRich Red preservative gt Expel entire contents of t
4. be made by applying fundamental cytological guidelines practiced by Cytotechnologists and Pathologists Optimizing Sample Adequacy gt Instruct health care provider on how to collect samples in the proper fixative and how to transport to the laboratory gt Follow manufacturer s recommended protocols for specimen collection and processing and use specified reagents and equipment gt Use only BD CytoRich Red or BD CytoRich Blue as the primary fixative for sample collection Other fixatives may impair sample adequacy gt Use only BD SurePath Precoat slides 1 The GYN application automatically loads when you start the BD PrepStain instrument workstation Exit this application to access a DOS prompt to display the Non GYN Version Check menu This screen provides access to all functions of the program gt If the workstation is running the GYN application select Quit to display the DOS prompt type in NONGYN and then press Enter The Non GYN Version Check menu is displayed gt If the workstation is already displaying the DOS prompt type in NONGYN and then press Enter The Non GYN Version Check menu is displayed 2 Select Run Version 2 80 from the menu and then press Enter This brings up the Non GYN Version 2 80 Main Menu The main menu provides access to all functions of the program 3 Select Sample Preparation and then press Enter The Number of Samples screen is displayed Note On this and all other BD Pre
5. choices a 8s 7 Vortex for 15 5 seconds and allow to sit for a minimum of 30 minutes F 8 Centrifuge for 10 minutes 600 g Program 3 9 Pour off supernatant fluid and vortex to homogenize sample See Step 13 for decanting instructions Il WASH Ge 7 IV PROCESS CAUTION Do not allow the slides to air dry prior to coverslipping For optimal results each settling chamber should be removed one at a time 10 12 13 14 15 16 17 18 19 20 21 22 If no visible or small pellet is identified add 6 mL of buffered DI H20 directly to 50 mL centrifuge tube vortex for 15 5 seconds and transfer entire contents to 12 ml tube If a moderate to large pellet is identified transfer a representative sample 1 5 drops to 12 mL tube and add 10 mL buffered DI H20 Transfer 12 mL tubes to centrifuge tube holder s and centrifuge for 5 minutes 600 g Program 4 Decant and vortex for 15 5 seconds to homogenize sample Decanting should be done in a single rapid motion by inverting each tube rack 180 degrees so as to not disturb the cell pellet While inverted carefully blot the mouth of all tubes in the rack with absorbent paper Turn the rack upright after 3 to 5 seconds Load the labeled 12 mL centrifuge tube holders onto the BD PrepStain Slide Processor for processing Place BD SurePath PreCoat slides on the slide racks in the same position as the tubes in the cen
6. protocol does not include using the BD PrepMate instrument however the laboratory can use its own judgment to validate other protocols gt How do split the non gyn sample if my cell pellet is too large How large is too large e If the cell pellet in the 50 ml tube is too large after decanting re suspend the pellet to homogenize the sample and transfer 1 5 pipette drops into the 12 mL tube e Too large is gt 1 mL gt Should spin down the non gyn sample before adding fixative e Yes if the sample is received fresh mix to homogenize then pour up to 50 mL of sample into a centrifuge tube Spin the sample down decant the supernatant and then add the BD CytoRich fixative as indicated in the protocol gt Can the non gyn sample be fixed in formalin e Formalin should not be used to fix any cytologic sample including gyn and non gyn samples gt How are mucoid samples processed e Mucus is considered a visible particle and needs to be removed from the sample prior to BD PrepStain processing e A variety of methods for reducing homogenizing or emulsifying mucus are available Examples include extended vortexing in a fixative that contains a mucolytic agent addition of mucolytic agents such as Mucolexx or DTT to dissolve the mucus etc e The laboratory can use its current methods for handling mucus when processing our non gyns e Your BD Diagnostics TriPath Representative can provide guidance at time of installation and
7. Helping all people live healthy lives Non Gynecological Applications Procedures amp Protocols TABLE OF CONTENTS Bibliography 18 Warnings and Precautions 19 Frequently Asked Questions 20 NOTES Required Materials Optional Materials gt 50 mL centrifuge tubes conical sterile or non sterile gt 30 mL pump dispenser for gallon container gt Urine type container leakproof sterile or non sterile gt Disposable transfer pipettes gt Buffered de ionized water pH 7 8 8 2 gt Squirt bottle 100 mL or 500 mL gt BD CytoRich Red preservative gt BD CytoRich Blue preservative gt BD PrepStain settling chambers gt 12 mL disposable centrifuge tubes gt BD SurePath PreCoat slides General Instructions gt When BD CytoRich Red amp BD CytoRich Blue are available directly use these for fixation of the specimen in the lab gt When there is no fixative available try to collect the specimen as fresh as possible and store them in refrigerated conditions 4 C Non Gynecologic Preservative Summary BD CytoRich Blue BD CytoRich Red Urine Bronchial Aspirates Washings Bladder Washing Body Cavity Fluids Sputum Brushings Fine Needle Aspirations For sputum urine and bladder washing s only use BD CytoRich Blue when the red blood cells need to be preserved for diagnostic reasons if their presence is not required use the BD CytoRich
8. No cloudy etc pamalray ajeq s eniu SJaMalAay juepunge ellued z ue p l 21 ulazold sAjod poojq uolzeundsqo ueuu g lej pouu Z Maf sdnoib gt 1 sou perq poob apene Z YOod ures poob abesare Z YOOd uolpeAsasaldg uBiu g a esapoud Z Mol Auen UAD UON 214070 ag poyujs JUIN UAS UON a UDI8O1 D ag poyujs 4U4 11N O u D uoN a UDI8O1 D qg poyujs 4U4 11N O u D uoN a UDI8O1 D qg poyujs 1u911n O u D uoN wPlyouD qg poyujs 4U4 311N O S UBWUWWOD uolpeindsqo Ayyjeno uleys UOIIBAJ S 1d Ayue n aD NOA uole1 idi lul pou adh uawidads ansoubeiq uone 11oD 21600A uoleued ld pue i quinN 3i souBeImq UOISS 22V e Arain S Walts AE Thomas P Bose S The anal Pap smear cytomorphology of squamous intraepithelial lesions CytoJournal 2005 2 1 4 e Bishop J MacFarlane K Cheuvront D et al Cell recovery and appearance in thin layer preparations in nongynecologic cytology Analyt Quant Cytol Histol 1998 20 229 237 e Bishop J Sims K Cellular morphometry in nongynecologic thin layer and filter cytologic specimens Analyt Quant Cytol Histol 1998 20 4 257 267 e Davis Devine S Day SJ Freund GG New red blood cell lysing fixative for use in fine needle aspiration and fluid cytology Acta Cytol 2003 47 4 630 636 e Gabriel C Achten R Drijoningen M Use of liquid ba
9. ample Removal of this material should be done according to current laboratory procedures e Recommendations on how to remove this material include pushing the material through a gauze or nylon mesh biopsy bag or using forceps to remove any clotted material gt How do collect fine needle aspirates FNA e Multiple options are available e Directly expel the FNA material into BD CytoRich Red e If air dried slides are required prepare direct smears first and place residual material directly into BD CytoRich Red e If conventionally prepared direct smears are preferred prepare these first and place residual material in BD CytoRich Red gt How do collect brush samples such as bronchial and gastric brushes There are multiple options to include e Deposit brush head in BD CytoRich Red Make sure there is enough BD CytoRich Red fixative in the specimen container to completely cover the brush during transport e Make direct smears first and then rinse or deposit brush head into BD CytoRich Red gt Can cerebral spinal fluids be processed with your method e Yes Simply follow the protocol provided e However since these samples may be limited in cellularity consider adding BD CytoRich Red to the collection fluid and letting sit for 30 minutes then pour into 12 mL BD centrifuge tube and centrifuge on program 3 decant vortex and place on BD PrepStain for processing gt Can you make a cell block with the BD Cy
10. ast three steps gt To confirm your entries and proceed select No and then press Enter 9 The Change Sample Stain Parameters prompt is displayed The default volume setting of Resuspension is always 600 and Sample always 200 Adapt these settings depending on the amount of cell pellet and number of slides that will be processed from each sample gt To change the sample or stain settings select Yes and then press Enter Refer to Change sample stain parameters on page 5 13 of the BD PrepStain Operator s Manual for details on how to make these adjustments gt To use the existing settings and proceed select No and then press Enter 10 The Tube amp Slide positions confirmed screen is displayed gt Confirm that the positions of the sample tubes and their corresponding slides match gt Select Yes and then press Enter The vacuum prompt is displayed and the alarm will sound Press any key to silence the alarm Turn on the vacuum pump wait a few minutes for it to warm up adjust pressure to 8 10 in Hg then press any key to continue 13 The Prime ALL Tubing prompt will appear gt If this is the first run of the day select Yes and then press Enter to prime the system tubing The system initializes and the pumps dispense reagents through the quad arm tubing and into the waste station gt For subsequent runs during the next eight hours select No then press Enter to skip the full priming function gt B
11. dark oo Explanations Solutions DI water was not used for BD PrepStain processing Do not use tap water pH of DI water was not appropriate Adjust the pH to 7 0 8 4 Staining intensity may be affected by ambient temperature Add extra water and or alcohol rinses to adjust stain to user preference See Operator s Manual for instructions on how to change stain parameters An inappropriate Hematoxylin stain was used Use Hematoxylin 0 5 provided by BD Diagnostics TriPath for non gynecologic specimens An inappropriate alcohol blend was used during processing Use BD PrepStain Alcohol Blend Rinse or prepare according to the following formula using reagent grade alcohol and isopropanol ALCOHOL BLEND RINSE Mix equal parts of 100 Reagent Alcohol denatured ethanol or 100 Ethanol AND 100 2 propanol isopropyl alcohol It is important that all glassware is clean and reagents are mixed in the correct proportions Additional troubleshooting guidance is provided in Chapter 7 of the BD PrepStain Operator s Manual If problem persists please contact Technical Support Test Method to Be Evaluated Non Gynecologic Specimen preparation using the BD CytoRich non gyn method BD Diagnostics TriPath Zaventem Belgium Objective To demonstrate equivalency between non gyn specimens processed using the BD CytoRich non gyn method compared to current methods used by the laboratory to include th
12. e Cytospin process direct preparations filter technique pick and smear etc Scope of Study 1 Determine time frame and approximate number of samples to be included prior to initiating the evaluation study The laboratory determines the total number of samples included in the study 2 Include representative samples from each general specimen category General categories might include respiratory tract urinary tract body cavity fluids deep seated vs superficial fine needle aspiration etc 3 Include when possible bloody mucoid serous and proteinaceous specimens 4 Include when possible normal and malignant specimens Split Sample Study Design Recommendations 1 Moderate to large volume fluid samples Mix sample well to homogenize Divide into two equal volumes Use current laboratory protocol to process the first aliquot and the BD CytoRich non gyn protocol to process the second aliquot 2 Small volume samples e g csf a Process fluid sample using current laboratory protocol Rinse residual material in BD CytoRich preservative and process using the BD CytoRich non gyn method Refer to Specimen Collection amp Handling section for type of preservative b Mix well to homogenize and divide into two equal volumes 3 Fine needle aspirates a Perform fine needle aspirate using current laboratory protocol Rinse needle tract in BD CytoRich Red preservative b Dedicate at least one fine needle aspirate pas
13. each slide rack is completed remove it within 10 minutes from the BD PrepStain instrument and decant the alcohol from the settling chambers into an appropriate receptacle and blot Rinse the slides one at a time Rinsing steps Take off the settling chamber from the first slide keep the slide in upright position above a waste bin Use a squirt bottle to flush loose cells and excessive liquids from the slide by spraying for 2 3 sec a stream of 100 alcohol just above the cell circle Rinse the slide in 100 alcohol and rinse twice in xylene or xylene substitute to clear Go through step 1 3 for all following slides Coverslip the slides according lab protocol Caution Leaving stained samples in alcohol for an extended length of time can cause the cells to de stain 20 When the BD PrepStain instrument finishes processing the Sample Preparation Complete prompt is displayed and an alarm sounds Press any key to silence alarm and continue 21 When you complete a Sample preparation you can either run another batch clean up the instrument or exit to DOS so you can start one of the other BD PrepStain applications For details on processing multiple slides for each specimen refer to Appendix C of the BD PrepStain Operator s Manual Observation Scant Cellularity Possible Explanations Solutions 1 The sample itself may be low in cellularity Concentrate multiple volumes by centrifugation combine the pellets and
14. efore each run a single syringe volume is automatically pumped through the tubing to ensure that the system is filled 14 When the priming cycle is complete the Is the PSSP tubing primed prompt is displayed gt To repeat the priming sequence select No and then press Enter gt To proceed with slide preparation and staining select Yes and then press Enter The DiTi dispenses buffered water into each centrifuge tube to re suspend the cell pellet Next the DiTi picks up a disposable tip and then a sample of the cell suspension is aspirated carried to its corresponding slide and deposited into the settling chamber 16 After a sample is transferred the BD PrepStain instrument dispenses an additional amount of buffered water When samples have been transferred to all racks the instrument pauses for ten minutes to allow the cells to sediment onto the slide 17 When the sedimentation pause is complete an alarm alerts the operator that the arm is about to move Staining is performed one slide rack at a time During each staining cycle each slide is pre washed in the appropriate reagent buffered water for Hematoxylin alcohol for EA OG then stained After the staining is complete the slide is washed with alcohol Each settling chamber is completely emptied between stains and washes 18 When all the slides on a rack have been stained the BD PrepStain system sounds an alarm then continues to stain the next slide rack 19 As
15. f azide For further information refer to the guidelines provided by the Centers for Disease Control Additional Warnings It is expected that good laboratory practices will be followed 1 Avoid splashing or generating aerosols 2 Centrifugation and fixation times or temperatures other than those specified may give erroneous results 3 Microbial contamination of reagents may give incorrect results 4 All reagents are stable until stated expiration date on the label provided recommended storage conditions are followed and maintained General Precautions 1 Do not pipette with mouth 2 Do not allow any reagents to come in contact with open wounds Precautions for Diagnostic Applications For all diagnostic and clinical applications of the BD PrepStain Slide Processor it is essential that the user strictly follow all instructions The user must ensure that requirements of the diagnostic protocol are carefully observed The BD PrepStain Slide Processor requires accurate positioning of all reagents samples and racks on the instruments workable by the operator The user should verify that all components are correctly positioned on the BD PrepStain Slide Processor and that the stain and solvent reservoirs are adequately filled before executing any program gt Can process BD SurePath and BD CytoRich slides at the same time e The BD CytoRich software program is different from the BD SurePath software progra
16. he Gyn program is in operation e A BD Diagnostics TriPath Clinical Applications Specialist can provide consultation on workflow issues such as handling a stat specimen gt Do I have to do a non gyn evaluation when converting to the BD CytoRich non gyn method e This depends on the requirements of your local regulatory authorities Please check local laboratory guidelines or contact your local BD Diagnostics TriPath Representative for more details gt Is there a recommended evaluation protocol e Although the laboratory is ultimately responsible for the evaluation protocol and study results documentation BD Diagnostics TriPath can provide consultation and recommendations gt How much BD CytoRich fixative is needed for the sample e For processing up to 30 mL in a 50 mL tube and up to 10 mL in the 12 mL tube e For fresh samples the general rule is a 1 1 ratio gt How long does the non gyn sample need to be in the BD CytoRich Red or Blue fixative before processing e A minimum of 30 minutes gt Can special stains be run on the BD CytoRich non gyn slides e Unstained slides can be prepared using the BD CytoRich non gyn method The laboratory would then be responsible for determining the staining protocols and performance for additional special stains gt Do I need to use the density gradient step BD PrepMate instrument for BD CytoRich non gyn processing e The current BD CytoRich non gyn
17. he needle aspirate directly into 10 mL of BD CytoRich Red preservative BD CytoRich non gyn processing procedure gt If fresh unfixed Follow General Procedure Step I IV Concentrate Fix Wash Process gt If received fixed Follow General Procedure Steps IV If sample is received fixed in BD CytoRich Red skip Step ll Processing Steps CONCENTRATION 1 Mix to homogenize the unfixed sample This can be accomplished by simple inversion or alternatively vortexing when a collection device is submitted i e brush sample 2 Assess for and remove clots particles and mucus 3 Transfer a 50 mL aliquot or for small volume samples the entire amount to a centrifuge tube g 4 Centrifuge for 10 minutes 600 g Program 3 9 5 Pour off supernatant fluid It is important to remove any visible clots or particulate matter Failure to do so may cause clogs in the BD PrepStain vacuum bundles Filtering may be accomplished by pouring the specimen through any coarse medium such as gauze Any captured material may be used for cell block procedures Follow standard laboratory procedures to homogenize and or emulsify mucous FIXAT ON This step may be omitted for prefixed samples Vortex sample for 15 5 seconds and proceed to step 10 6 Depending on cell pellet volume add 5 10 mL of BD CytoRich Red or Blue preservative Refer to collection procedures on page 3 for preservative
18. m Different sample and stain dilutions are associated with the BD CytoRich program In addition a different concentration of Hematoxylin stain is used for BD CytoRich non gyn vs BD SurePath test Therefore although BD SurePath and BD CytoRich specimens can be run on the same BD PrepStain Slide Processor they cannot be run at the same time gt Can substitute my own microscopic slides for the BD SurePath PreCoat slides e For optimal results use BD SurePath PreCoat slides provided which are coated with a proprietary formula enhancing cell adherence e The use of substitute slides is not recommended gt Can coat my own slides with a different coating agent such as Poly lysine e For optimal results use only BD SurePath PreCoat slides gt Can use commercially available pre coated slides e For optimal results use only BD SurePath PreCoat slides gt Can skip the water wash step e The water wash step removes any soluble proteins present in the sample If left in the sample these proteins will compete for binding sites with cells on the coated slides Washing the sample with water removes the proteins and enhances overall cellularity gt What do do if a body cavity fluid grossly looks like jello after sitting 30 minutes in BD CytoRich Red preservative e This type of specimen would be classified as having visible particles The jello like material should be removed from the s
19. pStain system screens you can use either the Tab or Enter keys to move the cursor navigate from one field to the next 4 In the first field type in the number of samples to be processed and then press Enter The number of samples must be a multiple of four Note If the number of slides to be processed is not divisible by four type in the next higher multiple of four and then add blank slides settling chambers and tubes to the slide tray to make up for the difference For e g if processing 6 specimens enter 8 in the first field and ensure 2 blank slides settling chambers and tubes are inserted into the blank positions 5 Go to the next field and type in the number of slides you want created for each sample For example you might want to create three slides from each sample centrifuge tube Details on processing multiple slides are illustrated in the BD PrepStain Operator s Manual Appendix C 6 Navigate to the next field and type in the number of slides that you want stained for each sample For example if you were creating 3 slides for each sample you might only want one of them stained using the current sample and stain settings 7 Navigate to the next field and either press Enter to confirm that the Next Tip Position is correct or type in the correct tip number and then press Enter The Reenter Run Information prompt is displayed gt To change any of your entries select Yes press Enter and then repeat the l
20. re process sample The supernatant of the concentrated specimen was not properly decanted Decant by inverting the tube or tube rack 180 degrees and blot to remove residual supernatant before returning the tube rack to an upright position Specimen was not vortexed prior to loading on BD PrepStain Slide Processor Vortex specimen prior to loading on the BD PrepStain Slide Processor to disperse and mix cell pellet Specimens fixed in BD CytoRich Red were not washed with DI water prior to processing Observation There is a halo or circle of stained cells on the prepared slide This may appear visually as a donut effect Possible Explanations Solutions 1 The wrong glass microscope slides were used Use the BD SurePath PreCoat slides The thickness of a BD SurePath PreCoat slide is optimized for the BD PrepStain Slide Processor and ensures a tight seal of the settling chamber to the slide Use of other slides may result in leakage around the O ring of the settling chamber resulting in a halo effect The vacuum pressure of the BD PrepStain Slide Processor may be too high Adjust the pressure to 8 10 psi Proteins in the sample are competing for sites on the coated slides Fix the sample in BD CytoRich Red and wash the fixed pellet with DI water prior to processing The Z max of the Quad may be too low Call Technical Support for guidance on troubleshooting Observation Staining is too
21. re stable for at least 30 days e Morphological stability has been demonstrated in specimens stored up to six months at room temperature e Specimens fixed in BD CytoRich Red Preservative Fluid can withstand shipment at temperatures ranging from about 5 to about 45 C gt Are BD CytoRich Red pre filled containers available e At this time BD Diagnostics TriPath does not provide pre filled containers for this purpose We provide bulk fixative and a commercially available pump can be purchased to facilitate filling specimen containers gt Does the BD PrepStain Slide Processor need to be placed under a laminar flow hood in order to process non gyn specimens e Since all specimens processed on the BD PrepStain Slide Processor are required to be fixed the processing does not need to be performed under a hood e However pre processing of fresh specimens should be performed according to the laboratory s protocol for safety and standard precautions gt Do you need a separate BD PrepStain Slide Processor just for processing Non Gyns e The BD PrepStain Slide Processor is capable of performing both gyn and non gyn applications on the same instrument e The total volume of gyn vs non gyn will determine the number of BD PrepStain Slide Processor s required in the laboratory Please contact your local BD Diagnostics TriPath Representative for more details gt Can prepare a stat non gyn specimen s if t
22. s for collection directly into BD CytoRich Red preservative 4 Brush samples Rinse or deposit brush head into BD CytoRich Red preservative after the smear is made If laboratory protocol requires the brush to be collected directly into fixative no smears made then mix the sample well to homogenize and divide into two equal volumes 5 Mucoid samples a For sputum processed using the pick and smear technique first prepare sample in the usual manner Fix remaining sputum sample in 30 mL of BD CytoRich Blue or BD CytoRich Red preservative if red blood cells may be removed from the specimen b For respiratory specimens processed using a homogenization process blending homogenize according to routine laboratory procedures Divide into two equal volumes 6 Fixed samples For best results the use of other alcohol based preservatives e g Saccomanno is not recommended Data Collection and Comparing Results 1 Determine the characteristics for each method to be compared such as cellularity preservation number of abnormal cells morphology etc 2 Compare using tools provided in this document 3 Derive conclusions based on comparative data analysis Frequently asked questions that may arise during implementation process are addressed on pages 22 25 Gross Data Specimen Fresh or Fixed Appearance Processed for Type of If fixed indicate Mucoid Bloody clear split sample Specimen type of fixative Yes or
23. sed cytology in serous fluids a comparison with conventional cytopreparatory techniques Acta Cytol 2004 48 6 825 835 e Hayama FH Motta AC Silva AP Migliari DA Liquid based preparations versus conventional cytology specimen adequacy and diagnostic agreement in oral lesions Med Oral Patol Oral Cir Bucal 2005 10 2 115 122 e Johnson T Maksem JA Beisheim BI Roose EB Klock LA Eatwell Liquid based cervical cell collection with brushes and wooden spatulas demonstrating a cost effective alternative monolayer slide preparation method Diagn Cytopathol 2000 22 2 86 91 e Kujan 0 Desai M Sargent A Bailey A Turner A Sloan P Potential applications of oral brush cytology with liquid based technology Results from a cohort of normal oral mucosa Oral Oncol 2006 42 810 818 e Maksem J Knesel E Liquid fixation of endometrial brush cytology ensures a well preserved representative cell sample with frequent tissue correlation Diagn Cytopathol 1996 14 367 373 e Maksem J Sager F Bender R Endometrial collection and interpretation using the TaoBrush and the CytoRich fixative system a feasibility study Diagn Cytopathol 1997 17 339 346 e Michael CW McConnel J Pecott J et al Comparison of ThinPrep and TriPath PREP liquid based preparations in nongynecologic specimens a pilot study Diagn Cytopathol 2002 25 3 177 184 e Motherby H Nicklaus S Berg A et al Semiautomated monolayer preparation of bronchial secretions using AutoCy
24. te PREP Acta Cytol 1999 43 1 47 57 e Nicol TL Kelly D Reynolds DI Comparison of TriPath thin layer technology with conventional methods on nongynecologic specimens Acta Cytol 2000 44 4 567 575 e Veneti S Daskalopoulou D Servoudis S Papasotiriou E loannidou Mouuzaka L Liquid based cytology in breast fine needle aspiration Comparison with the conventional smear Acta Cytol 2003 47 2 188 192 e Walts AE Thomas P Bose S Anal cytology is there a role for reflex HPV DNA testing Diagn Cytopathol 2005 33 3 152 156 e Weidman J Chaubal A Bibbo M Cellular fixation a study of CytoRich Red and CytoSpin collection fluid Acta Cytol 1997 41 1 182 187 e Weidmann J King L Bibbo M Modification of CytoRich Red fixative system for use on bloody Pap and fine needle aspiration smears Diagn Cytopathol 1999 20 2 95 98 e Wright RG Halford JA Evaluation of thin layer methods in urine cytology Cytopathol 2001 12 5 306 313 e Yang GC Wan LS Papellas J Waisman J Compact cell blocks Use for body fluids fine needle aspirations and endometrial brush biopsies Acta Cytol 1998 42 3 703 706 e Zardawi IM Duncan J Evaluation of a centrifuge method and thin layer preparation in urine cytology Acta Cytol 2003 47 6 1038 1042 Caution Some reagents contain sodium azide Sodium azide may react with lead or copper plumbing to form high explosive metal azides On disposal flush a large volume of water to prevent building up o
25. toRich collection material e Yes Any residual material can be cell blocked according to your laboratory s standard protocol gt Can you make unstained wet fixed slides with the BD CytoRich non gyn method e Yes refer to operating instructions gt Can you air dry a slide and stain it with Diff Quik Romanoffsky or Giemsa after the sample is fixed in BD CytoRich non gyn e It is best to prepare air dried slides with unfixed or fresh samples gt Can you perform Immunocytochemical ICC stains with the BD CytoRich non gyn method e BD Diagnostics does not have any data on ICC staining with non gyn cytology however the laboratory can evaluate this procedure as they normally would for other non gyn methods gt How do you store the centrifuge tube containing the residual specimen after processing e Follow your current routine laboratory procedure for storage and disposal of processed non gyn specimens e Note residual samples processed on the BD PrepStain Slide Processor are suspended in water To preserve cellular material consider adding BD CytoRich preservative for storage gt Are there any storage requirements for BD CytoRich Preservatives e Store at room temperature 15 30 C e Keep in a tightly closed and labeled container in a cool well ventilated area e BD CytoRich Red may be used up to the expiration date printed on the label e Specimens fixed in BD CytoRich Red preservative fluid a
26. trifuge tube holder If more than one slide is to be prepared from each sample tube follow the Non Gyn Slide Processing instructions in the BD PrepStain Operator s Manual for proper slide positioning Check levels of all reagent bottles For non gyn samples processing the use of BD PrepStain 0 5 Hematoxylin stain is recommended Place each labeled intake tubing into its corresponding reagent bottle Start slide preparation run Refer to the BD PrepStain Operator s Manual for software operation Remove each slide rack from BD PrepStain Slide Processor as it is completed Invert the slide rack and blot to remove any excess alcohol from the settling chambers Turn the rack right side up Taking one slide at a time remove and discard the settling chamber Rinse each slide one at a time first by using a squirt bottle Spray a stream of alcohol just above the cell circle then rinse the slide through a bath with 100 alcohol Clear with xylene or xylene substitute and coverslip according to laboratory protocol e Leaving stained samples in alcohol for an extended period of time can cause the cells to destain Slide Results and Cytologic Interpretation The BD PrepStain slide processor produces a uniform thin layer of cells in a 13 mm diameter circle The non gyn application of the BD PrepStain software allows for stained or unstained slides The preparations may contain single cells and cell clusters Interpretations should
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