Basics of #flowcytometry tweetorial!
As promised @DrGeeONE @DrPayelMondal @siti_richardson @nusrat_xahra @seyma_buyucek @Gagandeepk5MD @RubyMughal2 @anjuthevirgo @anueru432 @kis_lorand @AlanPath @Pathology36
1/11
#FlowCytomtery #HemePath #Flow #PathTwitter
As promised @DrGeeONE @DrPayelMondal @siti_richardson @nusrat_xahra @seyma_buyucek @Gagandeepk5MD @RubyMughal2 @anjuthevirgo @anueru432 @kis_lorand @AlanPath @Pathology36
1/11
#FlowCytomtery #HemePath #Flow #PathTwitter
What is a Flow Cytometry?
Flow= fluid
Cyto= cells
Metry =measurement
Flow cytometry is a process of performing measurements on cells (or particles) that are in liquid suspension-multiple characteristics gets measured simultaneously as cell passes through light beam 2/11
Flow= fluid
Cyto= cells
Metry =measurement
Flow cytometry is a process of performing measurements on cells (or particles) that are in liquid suspension-multiple characteristics gets measured simultaneously as cell passes through light beam 2/11
A Brief History of Flow Cytometry:
The first invention of a coulter counter was in 1949 by Wallace H. Coulter- method for detecting and counting particles suspended in a fluid medium
Later named the Coulter principle.
3/11
#beckmancoulter
https://www.beckman.com/about-us/company-history/wallace-coulter
The first invention of a coulter counter was in 1949 by Wallace H. Coulter- method for detecting and counting particles suspended in a fluid medium
Later named the Coulter principle.
3/11
#beckmancoulter
https://www.beckman.com/about-us/company-history/wallace-coulter
The First Coulter Counter
This is how simple it looked
Coulter was able to modulate the current by forcing a particle-filled fluid through a small aperture in an electrical current- allowing to count the number of particles in the fluid!
4/11
#flowcytomtery
This is how simple it looked
Coulter was able to modulate the current by forcing a particle-filled fluid through a small aperture in an electrical current- allowing to count the number of particles in the fluid!
4/11
#flowcytomtery
The two major achievements here were:
1.Reduced statistical error (by counting many cell)
2.Improved reliability of counter cells (as this showed that accurate cell counts were possible while revealing a lot of information regarding the patient sample).
5/11
1.Reduced statistical error (by counting many cell)
2.Improved reliability of counter cells (as this showed that accurate cell counts were possible while revealing a lot of information regarding the patient sample).
5/11
Early Models!
6/11
Source: JP, R. (2013). Wallace H. Coulter: Decades of invention and discovery. Cytometry A, 83, 424-438.
6/11
Source: JP, R. (2013). Wallace H. Coulter: Decades of invention and discovery. Cytometry A, 83, 424-438.
Pic 1: This was the first commercial Coulter instrument sold in 1956. and over 3,000 of these were manufactured- many directly in Coulter's basement in Chicago.
7/11
7/11
Pic 2: Model FN – released in 1968 – only the 4th iteration of the Coulter counter. This was the transistorized version of the Model B Coulter counter. This instrument has 5 digitron vacuum‐tubes which can display the digits 0‐9, giving a maximum count of 99,999.
8/11
8/11
What is Measured?
Bone Marrow image https://www.ncbi.nlm.nih.gov/books/NBK66050.1/figure/CDR0000258005__182/?report=objectonly
LN image https://my.clevelandclinic.org/health/diseases/15219-swollen-lymph-nodes
CSF image https://my.clevelandclinic.org/health/diseases/16854-cerebrospinal-fluid-csf-leak
#flowcytomtery #PathTwitter
9/11
Bone Marrow image https://www.ncbi.nlm.nih.gov/books/NBK66050.1/figure/CDR0000258005__182/?report=objectonly
LN image https://my.clevelandclinic.org/health/diseases/15219-swollen-lymph-nodes
CSF image https://my.clevelandclinic.org/health/diseases/16854-cerebrospinal-fluid-csf-leak
#flowcytomtery #PathTwitter
9/11
Solid tissue must be disaggregated before analysis
Broggi, A., Cigni, C., Zanoni, I., & Granucci, F. (2016). Preparation of single-cell suspensions for cytofluorimetric analysis from different mouse skin regions. JoVE (Journal of Visualized Experiments), (110), e5258910/11
Broggi, A., Cigni, C., Zanoni, I., & Granucci, F. (2016). Preparation of single-cell suspensions for cytofluorimetric analysis from different mouse skin regions. JoVE (Journal of Visualized Experiments), (110), e5258910/11
Next- will cover Optics of Flow Cytometry
To Be Continued....
To Be Continued....
This was the reason it took me so long to prepare the promised basic flow Tweetorial - 6 mon + Inter-professional Collaboration and entrepreneurship courses. And then Covid happened too! I am glad they are successfully over! ( okay what’s next )