Pipette Calibration System Student User Manual
Contents
1. Options H Help start I x ml i O Quantity one 4 6 7 lic Microsoft PowerPoint 7 10 04 am x Quantity One 4 6 7 E File Edit View Image Lane Band Match Volume Analysis Reports Window Help 1 aMtomation Manager EN _ TS Counting Colony Scanner Select Analysis Colon Counting Colony 7aunting ESI E ERU R X ray film 665 Open Display 0 2 ZoomB m oom Box VE Gispla 3 Transform 1 17 Ready AgarosAg6 Plus 14 Volume Rect Tool ef 5 V alume Free hand Tool b Vnlume Contour Toal Eal f Select Tool Print Image Volume Analysis Report le D D be D E D E E keyboard commands Chreclick copy a volume Shift click rotate a volume box Select Analysis Select Colony Counting from Drop Down Menu Q cqui Stop Options Save After Scan Hide Grid al 1 13 15 17 dd 21 23 25 ze 29 Highlight Saturated Pixels Options H Help start PN x fa E Quantity one 4 6 7 7 00 v P 10 04 am lic Microsoft PowerPoint Counting Colony Adjust Area of Colony Step 1 Define Region and Count e For best results adjust2. Define Counting Region Drag the cursor from center to edge of dish image Count Sensitivity ERI f l Averaging 1 3 5 7 Plaque Step 2 Adjust Results White Blue Colony count 0 75 a 5 White and Blue Colonies Adjusted coun If you know you have white and blue colonies in the image and there are two clear peaks on the histogram to the right ofthe background peak you can use the histogram to distinguish between these types of colonies Count vs Peak density 111 Results Vw ita Bius Colony count s3 152 Adjusted count 152 Cutoff White Blue _ White peak _Blue peak Step 3 Tools options Count ws Peak density T Ignore region x Show data area x Mark white colonies Make Colony Cutott __ whiterbiue xj Mark blue colonies Totals T division mark amp Erase Colony White 0 tae A Blue 75 Step 4 Save To Batch File Batch mode New batch batch Save Count Load Next Batch File Becky GFP Neg 03 batchcount 1 xls Fig o6 Using the Vvhite bBlue slider Drag the White Blue slider to the left until itis positioned between the two peaks The white colony data range is indicated by gold on the bar beneath the histogram and the blue colony data range 15 marked with blue Count Name Operator 2010 07 09 lOhr O3min 10 06 46 Coun
3. i Colony Colony Counting using Darkfield BioRad UMAX GS 800 Scanner BioRad TC10 Cell Counter Viewers and Bio Rad Quality One Software With Thermal Printer Counting Cell and Colony SOP and Description Prepared by Bob Morrison SILCC CPLS Instrumentation Specialist STLCC CPLS Morrison 9 30 2011 August 2008 Initial Latest Rev Mar 11 Page 1 Counting Cell Counter Bio Rad TC10 TC10 Automated Cell Counter 145 0001 100 240 V includes instrument USB key 30 TC10 dual chamber counting slides 60 counts 1 5 TC10 trypan blue dye Slides 145 0011 30 dual sides per box 60 total counts Counting time 30 sec Cell concentration range 5 x 10 1 x 10 cells ml Cell diameter range 6 50 um Sample volume 10 ul Data storage 100 counts Data export Via USB flash drive Printer connectivity Yes Dilution calculator Yes Image of cells available Yes Dimensions W x D x H 19 x 15 x 25 4 cm 7 5 x 6 x 10 Weight 2 2 kg 4 8 Ib without the external power supply 05 Export to Flashdrive Y 4 On Off The TC10 cell counter is an automated device that provides a total count of mammalian cells and a live dead ratio in one simple step with accurate reproducible results Count cells prior to cell culture or before starting processes and analytical techniques that require an accurate and consistent number of input cells The TC10 automated cell counter helps you avoid the tedious task of counting c
4. Counting Colony Viewers Counters at FV Key Features Even glare free illumination Light is spread uniformly over the entire culture plate Colonies are bright and easily distinguished Adjustable dish holder for centering both round dishes and square culture plates Adjustable focusing rod Lens rotates a full 360 Built in tilt leg Optional 1 5X auxiliary lens fits over standard lens increasing magnification to 3X Footprint 12 5in wide x 14 in high x 12 in depth Quebec Darkfield Model 3330 1 An adjustable dish holder for centering round dishes with diameters up to 100mm and square culture plates up to 100mm x 100mm 2 An optional 1 5X auxiliary lens fits over the standard lens increasing magnification to 3 0X 3 The adjustable focusing rod allows the 1 5X standard lens to be raised or lowered The lens also rotates a full 360 for ready access to culture plates 4 A built in tilt leg may be mounted in the front or rear of the instrument allowing a convenient tilt angle or it may be locked flat to the instrument base 9 A white ruled Wolffheugel counting plate is included 6 Internal standard light bulb 110V plug connection On Off switch Quebec Darkfield Model 3325 3327 STLCC CPLS Morrison 9 30 201 1 Page 20 Counter Automated Invitrogen Countess The Countess Automated Cell Counter uses trypan blue staining combined with a sophisticated image analysis algorithm to produce accurate cell
5. MIT HHE HE E E m Em E pz S elect Y resolution amp 3 5 Image size 4 17 Mb Select image and then Photograph Change Light from Reflective to Transmissive to best see your specimens Preview Scan and wait for image outline to appear Adjust crop box to contain just your specimen 5 Select ACQUIRE to capture the image Hit STOP if image preview fails to appear Q cqui t Stop Options Save After Scan mes Make Backup Highlight Saturated Options H Help Quantity One 4 5 7 9 5 e RE 10 03 am me Quant Counting Colony Scanner Examine Acquired Image Step Select Application lick Select FELEEELEELEEEESS pepe ER RR RN film D 3 pen _ me ae rer iat lee 2 Zoom Transform Volume Rect Tool Volume Free hand Tool Volume Contour T aal Select Tool Print Image M Le D D Le b D D D E Volume Analysis Report Keyboard commands Ctrl click copy a volume Shift click rotate a volume box Q cqui Stop Options Save After Scan Hide Grid gt iMakeBackup Copy 43 15 17 18 4 23 25 27 29 Highlight Saturated Pixels
6. court screen Use the up ar dewn keys and press Enter to access the following screens Dilution caleulatar View image Print count and View histogram Helpful tips foreach green are provided below Caluten Calculator 1 In the eakulstar the current court used as the starting cell concentration The that ready ta be is highlghted Press the upar down arrow key te find the corect value then press Enter te confirm the 2 print the Dilution caleulatorsereen press the dawn 5 Counter Cell Counting Bio Rad Quickguide Pg 2 2 Fig 3 Gure mt count sc mee nshowing a total cell count without torpa n blue due TE10 Automated Cell Counter Current count Live count 2 28 x 10 cells ml FA we cells 70 10 12 55 PM February 20 20 Sec Dilution calculator Fig 4 Gurrentcountzcrme nshowing total cellcount Ime cell count and percentage of lie celk cell viability measured via trypan blue dye exclusion View mags Use the up arrow key to zoom in onan image and samire eels in detail Use the down key te moarn aut Print Count The autarnmted cell counter allows you to print the cell count Wingy the TOTO thermal label printer 1445 00608 connected ta USE part Aor USB part Boon the cell counter View His gram SIECC CPLS Morrison 9 30 2011 5 Counter TC10 Exporting Images of Cells Ex
7. includes TC10 verification slide protocol AE Price approx 360 retail du ME p SN gQ Result of Verification Test March 28 2011 8 41 System verification performed Side A Pass 3886 of 3886 0 50 Side B Pass 195 of 195 0 50 Rerun 9 30 11 same results as above From the Home screen select Options then Enter 2 From the Options screen select System Test and Enter 3 Follow instructions to insert side A of the verification slide then side B then a blank read is made 4 Results of the test are displayed and can be exported to a flashdrive using the Export Previous counts function 5 Complete instructions on the verification slide protocol can be found at the hotlink shown below Hot Link to Bio Rad TC10 Verification slide protocol 2 pgs STLCC CPLS Morrison 9 30 201 1 Page 8 Counting Bio Rad TC 10 Printer Thermal Bio Rad Part 145 0005 TC10 Thermal Label Printer includes label printer USB cable 1 roll of 185 labels 145 0007 Thermal Printer Labels 1 roll of 185 labels for TC10 thermal label printer USB connect to TC10 Counter i fom i Imm Manufacturer Citizen Manufacturer Part CT S281RSU WH P A Cost Central Item 11050383 Product Description Citizen CT S281 Label printer two color direct thermal Roll 2 3 in 203 dpi up to 189 inch min serial Printer Type Label printer direct thermal two color monochrome Approximate Dimens
8. mix the cells and dye 2 Within 10 minutes of mixing pipet 10 Lil of the mixture into tha outer opening of either of the two chambers of the courting slide Figure 1 Erf armine Call Gains VIOrTrIiSon 1 Press Home and insert slide 2 The T10 cel counter autarratically detects the presence af the side and initiates a count The Tig counter also automatically detects the presence of typan blue dye Important De not rermeve the side ar interrupt the Irstrurnent whike ts perferre the count a Far samples without trypan blue dye an the Curent count screen the counter provides the total well count per mli F eure 4 Far samples with trypan blue dye an the Current court screen the counter provides the total cell count per mil Ime cell count per ml and percentage of ne cells Figure 41 It abo accounts fara 1 1 with the dye and provides a dilution cal ulation based an Ive calg Important lf the number of cells is above or be bw the range detected by the Instrument a Value aut of range message appears an the Current count screen Press the dawn arw key ta select View image then press Enter View the image of the cells and determine whether the sample should be diluted careentrated Repeat the cell eaunt amp Ores the instrument completes the cell count renews the slde fimm the slde skt Additional Analysis Options ral analysis options are avaiable in the Curent
9. the Gel Doc zoom lens so that Define Counting Region the cursor from center to edge of dish image Count Sensitivity f l C1 5 7 Plaque Step 2 Adjust Results White Colony count 0 0 Adjusted coun Count vs Peak density Cutoff amp White Blue TEP 1 DEFINE REGION and Basic Count Controls 1 5 Step 3 Tools options 2 Select Icon to Define Region at Center of Image Ignore region xjShowdataarea 3 Drag cursor to move Blue diameter circle Make Cory x Mark white colonies 4 Adjust sensitivity higher to find more colonies Mark blue colonies SS Erase Colony 9 D Step 4 Save To Batch File Batch mode New batch Open batch Save Count Load Next so Va Batch File Becky GFP Neq 03_batchcount 1 xls Count Name Operator 2010 07 09 lOhr O3min l0 04 54 Count Comment Close Reset Help Start 9 f fx a 6f 23 Quantity One 4 6 7 Microsoft PowerPoint 9 2 SJ C 10 05 am Quantity One 4 6 7 m E igi x Counting Colony Adjust Count vs Density m gt Step 1 Define Region and Count 1 If there is a clear peak on the left end of the Define Counting Region e Diag the cortos hon Conr to edge of dish image YA colony histogram It Is probably due to background intensity or noise in the image 2 Use Cutoff slide to a
10. 8 Count Comment Use STEP 3 to Adjust items in out of the counts 2 Erase colony then pick items from image that are NOT part of the colonies 3 Make Colony then pick items to be added as colonies yellow markers start G fa E lw Quantity One 4 6 7 Microsoft PowerPoint 7 10 07 AM ES Counting Colony Save Count Data to File Excel other gt Step 1 Define Region and Count Ek For best results adjust the Gel Doc zoom lens so that Define Counting Region Drag the cursor from center to edge of dish image __ Sensitivity ait Averaging 471 6 3 6 5 7 Plaque Step z Adjust Results White Blue Colony court NEN Adjusted count n Count vs Peak density STEP 4 Select New Batch Enter or edit filename Ww Fite Blue Step 3 Toolsoptions Ignore region x Show data area x Mark white colonies x x Mark blue colonies Erase Colony White 152 Blue 0 Step 4 Save To Batch File x Batch mode Mew batch Open batch Save Count Load LT 7 Batch File Becky CFP Neg 03 batcheount 1 xls iJ amp Eu ale Count Mame Operator 2010 07 09 lO0hr 10 11 41 Count Comment Close Reset start E a x fa Et Quantity One 4 6 7 Microsoft PowerPoint 2 zi D 9 10 11 AM
11. and viability counts in just 30 seconds The algorithm also measures average cell size of live dead and total cells to give you all the data you need to proceed with your experiments The measurement range extends from 1 x 104 to 1 x 107 cells ml with an optimal range from 1 x 105 to 4 x 106 cells ml broader than that of a hemocytometer view technical notes for more comparison data The optimal cell size is between 5 um to 60 um view validated cell lines A handy dilution calculator even helps you determine how to prepare your sample for your next passage or experiment STLCC CPLS Morrison 9 30 201 1 Example Only Presently Not at FV IO 3 1 4 Counters Automated Cel Count Homeo tonsa easured concentration 2 10 Call suspension dilution cells ml The Countess Automated Cell Counter eliminates the tedium and subjectivity of manual cell counting Automated counting frees up your time reduces eye strain and minimizes subjective judgments that can lead to error It takes 3 simple steps 1 Mix 10 ul of sample with 10 ul of trypan blue and pipet into Countess chamber slide 2 Insert slide into the instrument 3 Press the Count cells button results are displayed in 30 seconds Page 21 Other Protocols or Notes e Future home of other or more details protocols STLCC CPLS Morrison 9 30 201 1 Page 22
12. djust per notes below Count Sensitivity f l Averaging 1 3 C5 7 Plaque Step 2 Adjust Results White Blue Colony count 0 75 Adjusted count r ae Count vs Peak density NN If background 15 being detected as colonies you can use the histogram and the Cutoff slider to correct this Cutoff Drag the Cutoff slider to the right until itis centered on the nght edge of the background peak Step 3 Tools options 7 Ignore region Show data area x Mark white colo k Make Colon d Mark blue colon Erase Colony Step 4 Save To Batch File Batch mode New batch Open batch Save Count Load Ne Batch File Becky GFP Neg 03 batchcount 1 xls Cutoff mark Count Name Operator 2010 07 09 lOhr O3min 10 06 46 Count Comment Fig 6 5 Using the cutoff slider The yellow portion ofthe bar beneath the histogram marks the range ofimage data has been designated as background noise andis not being considered for colony start 3 f amp a t e quantity one 4 6 7 451 G counting purposes The gold portion of the bar marks white colony datarange e ML m i 4 0 pis Quantity One 4 5 7 E 8 x E Counting Colony Adjust White vs Blue 4 Eh For best results adjust the G el Doc zoom lens so that Step 1 Define Region and Count oo o
13. ells with a microscope and hemocytometer which produce varying results The TC10 automated cell counter uses multifocal plane analysis to assess cell viability And it demonstrates accurate cell counts of viable cells and across a range of cell ST LCC CPLS Megi ee strates and cell sizes Page 2 Counting Cell Bio Rad TC10 Loading Slides Loading Slides Handle the TC10 counting slides using the edges and avoid touching the optical surface of the slides Important hen loading the sample place the pipet tip at a 45 deqree angle at the bottom of the sample loading area halt circle at outer end of the chambers slide thetip along the surface and carefully touch the apex of the halt circle Figure Once the tip is stopped depress the plunger to begin the capillary loading process Care should be taken to avoid visible bubble formation or back splatter Do not overfill or undertill the chamber Load here Fig 7 Loading the C10 counting slide Important Load samples only into the outer ends of the counting chambers Figure 9 he center areas are open to promote uniform distribution in the counting chamber Do not use the center areas for sample loading to avoid Load here an undercounting of the cells in the sample Fig 8 Correct and incorrect loading are C10 cell counting slides cannot bs reused Dispose of used slides as biohazardous was according to your local environmental health ana safety regulation
14. ions WxDxH 4 2 in x 7 1 in x 4 1 in Approximate Weight 1 4 lbs Localization United States Max Media Size Standard Roll 2 3 in Print Soeed Up to 189 inch min max speed Max Resolution B amp W 203 dpi Interface Serial Barcodes Code 93 Code 39 QR code EAN JAN 8 EAN JAN 13 UPC A Codabar Code 128 PDF417 ITF Power AC 120 230 V Microsoft Certification Compatible with Windows 7 Hot Link to Bio Rad Citizen TC10 Printer Manual pdf 33 pgs STLCC CPLS Morrison 9 30 201 1 Page 9 Counting Colony Using GS800 Scanner Quality One Software NN Turn on Scanner and wait for both lights to show steady green this is the ready state Turn on Host computer logon as Operator SMET jy Select Quality One to scan acquire and image nternet C pue s expert JUantity One E 2 obe 10 1LUUU Y os JUantity zj Bit JL ESL Link to BioRad Quantity One 1 D Analysis Software Users Manual pdf STLCC CPLS Morrison 9 30 201 1 Page 10 Quantity One 4 5 7 File Edit View Tr Salt Co unting Colo ny Quality One Select Scanner 1 x ul Select Scanner s Open 12 oom et 3 Transform Volume Rect Tool 15 Valume Free hand Tool uantity One X ES 4 Volume Contour Tool E E emn Select Scanner Selec
15. porting the Image of Cells from the Current Count The image from the most recent count can be downloaded onto a USB flash drive and viewed on your computer in JPEG format export the image 1 Press Home to go to the Home screen 2 Use the down arrow key to go to Options and press Enter 3 Choose Export current image and press Enter 4 lf there is no USB flash drive inserted into the instrument Insert USB flash drive into Port A to export current image Press Home to cancel is displayed on the screen Insert a USB flash drive into the USB port A If a USB flash drive is already inserted into the USB port A the export starts automatically A bar snowing the progress of the image download is displayed the screen When the image export is complete File saved as CELLxxx JPG Press Home to exit is displayed 6 exit press Home f Remove the USB flash drive from the instrument Note If a new slide is inserted while the instrument is exporting an image the instrument queues the count until it completes the export The instrument then detects the slide and asks If it should start counting the sample in the new slide If a new slide is inserted after the image is exported the instrument starts counting the sample in the new slide SILCC CPLS Morrison 9 30 201 1 Counting Bio Rad TC10 Verification Kit Bio Rad part no 145 0014 Kit for validation of TC10 automated cell counter functionality 2
16. s VIOITISOTn Counting Cell Bio Rad TC10 User Interface IVIOMISON d User Interface he user interface contains a color LCD screen and the four navigation keys listed below Home for accessing the Home screen or for exiting any screen m Up and down arrow keys for moving to the desired setting within a screen m Enter for confirming a selection Home Screen The Home screen can be accessed from any screen by pressing Home This screen Figure 2 provides access to the following choices Count cells Count cell amp initiates a cell count When this option is selected the Instrument will automatically start counting if a slide already inserted If there is no slide in the instrument the Insert slide screen will be displayed Previous counts for viewing results from up to 100 previous counts stored in the instrument results are organized by date time stamps with the most recent count shown first Fig 2 Home screen Export previous counts for exporting results from the instrument onto a LI SB flash drive Dilution calculator calculates volume adjustments required to achieve the cell concentration needed for the next experiment Options for selecting additional operations Hot Link to Bio Rad TC10 User Manual pdf 36 pgs Counting Cell Counter Bio Rad QuickGuide pg 1 2 The TO10 automated cell counter counts mammalian cells in one simple step with or without tr
17. t Comment As you drag the slider the numbers of white and blue colomes will change in the dialog and in the text box on the image Also on the image you should see the marked white colonies gold triangles change to blue colonies blue squares ote Ifthe blue colonies are not marked on the image check to make sure that the Mark Blue Colonies checkbox at the bottom ofthe dialogis checked Start M x a E v o 65 Quantity One 4 6 7 c3 Microsoft PowerPoint Quantity One 4 6 7 File Edit 1 x Counting Colony Adjust counted items Ek For best results adjust the Gel Doc zoom lens z0 that gt Step 1 Define Region and Count Define Counting Region Drag the cursor from center to edge of dish image __ Sensitivity ait Averaging 471 6 3 6 5 7 Plaque Step 2 Adjust JTJ 1 White Blue Colony count 153 Marked Colonies Adjusted coun 77 Count ws Peak density lI ul Cutoff whiter Blue A Step 3 Taonolz aptiansz Ignore region x Show data area x Mark white colonies amp Make Colony Mark blue colonies Totals Erase Colony white 153 Blue Step 4 Save To Batch File _ Batch mode Mew batch Open batch Save Count Load Batch File Becky GFF Neg 03 batchecount l xls Count Hame operator 2010 07 09 lOhr O2min l0 07 3
18. t Tool Print Image 3 Gel Doc EQ 22 9 Volume Analysis Report a ChemiDoc EQ ChemiDoc XES 1 Choose Select Scanner on the T Volume Quick Guide menu 2 Select GS 800 as scanner type Fluor 8 Fluor S Fluor S Mane VersaDoc PMI FX JBst xX GP LS Mortisoric9 30 201 a Quantity One 4 6 7 ica Microsoft PowerPoint Quantity One 4 5 7 Counting Colony oet Light Preview Acquire magg G 2 x Volume Analysis Report Bottom 19 Aight 20 8 Step Select Application Quick ai i PERE EEE EEE EEE eet x PX E i a EEE Open 2 Gray filn No ES ER Hoe a 112 Zoom Box 2 2 l 9 13 Transform Filter Red Greer Blue M iE BEES I Light Reflective Thynsmissive 11 Volume Tool ERE h Volume Free hand Tool a Step Il Select Scan Area Bee ks i Valume Contour Tool a Previ M Preview scan BEEE a Click and drag in diagram to set scan ares Fr 7 Top Left 11 5 18 7 saa 81 a Keyboard commands Chreclick copy a volume Shift click rotate volume box Step 111 Select Resolution resolution E Ei E p i m lil NS
19. ypan blue staining and assesses cell viability via trypan blue dye exclusion Important Carefully read the safety information and instrument operating specifications provided in the T Automated Call Counter Instruction Manual before using Hot Link to Bio Rad ie system TC10 User Manual preparing Samples pdf 36 pgs The automated cell counter is designed to count suspended cells that have been grown as adherent or suspension cells The TC 10 counter demonstrates high reproducibility when counting cells within the ranges of Fig 1 Loading the sample onto a slide 5 x 104 1 x 10 calls ml and 6 50 um cell diameter Important Hande the T10 counting slides using the and avoid touching the optical surface of the slides counting slides cannot be reused Dispose of used slides as biohiazardous waste according to your environmental health and safety regulations Praparing amplas without TC10 Trypan Blua Dya Fipet 10 jl of the cell suspension into the outer opening of either of the two chambers of the counting slide Figure 1 Praparing amples with 10 Trypan Bluse Dye 1 In a micro test tube combine 10 plot the cell suspension with 10 pof T10 trypan blue vynen counting the sample in duplicate combine 20 pl of Fig 2 Inserting the slicle into the TOU cell counter the suspension with 20 ulot TG 10 trypan blue Genty pipet up and down ten times to
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