Showing posts with label flow cytometer. Show all posts
Showing posts with label flow cytometer. Show all posts

Friday, February 1, 2019

*** desktop flow cytometer and cell counter survey


LUNA-FL . $9.7K 

Moxi Go 2, VWR $21K
https://us.vwr.com/store/supplier/id/0000012763/ORFLO+TECHNOLOGIES

Guava Millipore

CytoFlex $76K ~ $110K. 


advice from others:
"You might also look into the LUNA-FL or Countess FLII systems if the end use is as described, they are basically automated low-power epifluorescent scopes.  There are disposable and reusable slide formats. These are instruments much cheaper in comparison.  However, the Countess might not be able to assess yeast viability.

I have a Countess that we use for simple stuff, and we can get rough phenotypes for abundant surface markers.

I cannot recommend the Muse because it is my understanding that it’s completely kit based.  The Guava has been reliable in my experience, but I haven’t used one in a few years. "



SpectraMax® MiniMax™ 300 Imaging Cytometer, Molecular Devices, $45K 




"I strongly recommend trying your yeast samples on a demo Countess II/II FL instrument before purchasing one.  Yeast cells are generally very small and may be below the size cut-off for accurate counting and viability.  The instrument will see and count any cells larger than 4 um in diameter, but will only give correct viability for cells larger than 7 um.  If you open the histogram option on the Countess II results screen, you can see the diameter range the instrument sees for the sample. Unfortunately, we do not have example data for yeast.  "

================
Main fluophores: 
  Propidum Iodide (PI), dihydroethydium (DHE), and dihydrorhodamine (DHR)

My understanding of the basic configuration are (based on CALIBUR):
 blue laser 488nm + red laser 635nm (optional)
Detector channels: FSC, SSC, FL1 green 530nm, FL2 yellow 585, FL3 red 670nm, with an optional  FL4 661nm. 

http://qa.ff.up.pt/~quimicafisica/405.pdf

Vendors and models

BioRad S3e Cell Sorter

easyCyte
http://www.emdmillipore.com/US/en/life-science-research/cell-analysis/guava-easycyte-flow-cytometers/zLWb.qB.7eAAAAE_1rFkifKv,nav?cid=BI-XX-BDS-P-GOOG-FLOW-B323-1017

http://www.emdmillipore.com/US/en/life-science-research/cell-analysis/guava-easycyte-flow-cytometers/specifications/7syb.qB.D6MAAAFBV7o7FnRb,nav;pgid=WZtMKfaYs81SRpEoh9VgbI720000CVPV_BBr;sid=PkPRmcIRQ1_emZUkxcfBESxEh8UTxgGbg-NYIv436iSRwUp_5oE3LgZPVeQAlXodKzH_gBFn_X4oMZfvewXUpohLXOJVkMcVd4OD_HWFx2Bh78hq5ZpOWn9X5RVu417NYgP_gBFnti0JWwEsx-uzOEYY

http://www.emdmillipore.com/Web-US-Site/en_CA/-/USD/ShowDocument-Pronet?id=201601.186

ACEA NovoCyte 1000 $40K ?
http://www.aceabio.com/products/novocyte/
Novote Cyte model 2000, 3000 have more options.

BeckmanFlow.com CytoFlex
http://beckmanflow.com/?page_id=379/?pi_ad_id=105930129160

Accuri C6PLUS  $49K

EmdMillipore Muse: $15K
http://www.emdmillipore.com/US/en/life-science-research/cell-analysis/muse-cell-analyzer/assays-for-muse-cell-analyzer/cell-health-assays-for-muse-cell-analyzer/2nab.qB.Jp0AAAFQ46hauq.k,nav#mch100102
Con: Closed system. Cannot be modified ?

Attune NxT Flow Cytomer

Stratedigm FLow Cytometer

http://www.ncbi.nlm.nih.gov/pubmed/16680701
http://www.biocompare.com/Editorial-Articles/167700-What-To-Look-For-in-a-Personal-Flow-Cytometer/



Sales rep from BD on Gallios and CytoFlex
As per my voice message, I understand you are looking for a quote for 6 color Gallios Flow Cytometer, but we now have a new, much smaller flow cytometer with greater capability than the Gallios, and less expensive.   It is called the CytoFLEX Flow Cytometer.   The CytoFLEX is available in 21 different configurations from a 1 laser/4 color system to a 3 laser/13 color system in a small box (16in x 16in x 13in).   It is completely upgradable to any configuration at any time.   A 96 well plate loader option is available as are 561 nm and 375 nm laser options.   Independent testing has proven this instrument to be one of the most sensitive instruments in the market today.
 If the 561 or 375 nm lasers interest you, please let me know and I will forward additional information on our CytoFLEX “S” System.

Monday, October 16, 2017

fluorescent labels



http://www.clontech.com/US/Products/Fluorescent_Proteins_and_Reporters/Fluorescent_Proteins/Fusions/Red

http://www.clontech.com/US/Products/Fluorescent_Proteins_and_Reporters/Fluorescent_Proteins/Fucci_Cell_Cycle_Vectors?sitex=10020:22372:US&PEBCL1=PecQzqNVe9wzSLzBdE2uoNDO4A&PEBCL1_pses=ZGA8D8F214383771F0C0DB76A3075304A12F9A86A5827CFE3228A1B0DF9BA9D13CA93EB3593F96367AD0B93113BDB1E664E8DA5C56C31220DE


Saturday, October 7, 2017

Moxi flow, next generation flow cytometer

Moxi flow ~ $9000
moxiflow kits
https://www.orflo.com/product_p/mxf002.htm
flow cytometer cassettee, 2 test per cassette, $160 for 25 cassettee and 50 test. This means about $3.2 per test.  For 100-day assay of 50 strains, it means $160*100 = $16K.

Moxi flow Go 2, $21K from VWR
https://us.vwr.com/store/supplier/id/0000012763/ORFLO+TECHNOLOGIES

next generation flow cytometer
https://www.ncbi.nlm.nih.gov/pubmed/28104919

https://www.nature.com/articles/nbt.1919.pdf


Wednesday, September 14, 2016

advantages of the 561-nm (Yellow-Green) Laser for the flow cytometry.


a brief list of advantages of the 561-nm (Yellow-Green) Laser for the flow cytometry.

all the RFPs, GFP, CFSE and Fitc when really bright bleed into the PE channel horribly when using just a 488 laser. If you are using the 561 for PE there will be almost no spillover.

—  tdTomato detection (fluorescent protein 283% brighter than eGFP) 
—  MitoTracker Orange (accumulation is dependent upon membrane potential and the dye is well-retained after aldehyde fixation)
— CRISPR-Cas9, an emerging gene-editing (“genome surgery”) tool that frequently uses mCherry reporters. And yellow-green laser is required for optimal excitation of this fluorescent protein
— up to 3.7-fold increase in the stain index with antibodies conjugated with PE and PE based tandem dyes 
— greatly expands the choice of fluorochromes

Sunday, August 16, 2015

flow cytometer training


http://www.thermofisher.com/us/en/home/support/tutorials.html?socid=social_20150816_50608416&adbid=1195673353783588&adbpl=fb&adbpr=183929358291331

bio125
bio233

Thursday, April 9, 2015

flow cytometer, sample probes tend be clogged by yeast cell clumps.

When yeast cells were too dense, the sample probes of Calibur tend to be clogged. Presumably because chained yeast cells clumps.


Thursday, March 26, 2015

*** Calibur, gating for wildtype and vector for bio125 MSH2 yeast strains

__ AGY 124 (vector control), AGY125(MSH2 control), grown in SD-His-Trp-Ura for 2 days
__ take 100ul to 2ml water
__ point sonication, level 2, 4 times

Calibur, E03 FSC, 952 SSC, choose gate R1 for yeast cells.

template file: tempalte20150320-R1R2-v2






Tuesday, July 9, 2013

DNA content, rDNA cluster size detection by flow cytometer

Marcus Nordberg used flow cytometer to detect genome size variation. He thought genome variation is primarily due to rDNA cluster size changes.  In yeast, rDNA cluster size -> translational speed and accuracy OR extra-chromosomal rDNA circles OR genomic instability.  rDNA and ribosomal function is related to lifespan. So, flow cytometer can be used to quickly phenotype rDNA sizes.

Alternatively, qPCR, short-reads mapping, and southern hybridization can be used. In this case, standardized rDNA clusters will be used to generate a standard curve. The rDNA cluster reference strains are available in  David Bedwell's lab at UAB






Wednesday, February 22, 2012