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\author{woozing}
\pdfinfo{
  /Title (ptc-c9-cell-counting-haemocytometer.pdf)
  /Creator (Cheatography)
  /Author (woozing)
  /Subject (PTC - C9 (Cell Counting - Haemocytometer) Cheat Sheet)
}

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        \hspace*{-6pt}\includegraphics[width=5.8cm]{/web/www.cheatography.com/public/images/cheatography_logo.pdf}}
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    \vspace{-2pt}\large{\bf{\textcolor{DarkBackground}{\textrm{PTC - C9 (Cell Counting - Haemocytometer) Cheat Sheet}}}} \\
    \normalsize{by \textcolor{DarkBackground}{woozing} via \textcolor{DarkBackground}{\uline{cheatography.com/146689/cs/31813/}}}
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  \mymulticolumn{2}{p{5.377cm}}{\bf\textcolor{white}{Cheatographer}}  \\
  \vspace{-2pt}woozing \\
  \uline{cheatography.com/woozing} \\
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  \mymulticolumn{1}{p{5.377cm}}{\bf\textcolor{white}{Cheat Sheet}}  \\
   \vspace{-2pt}Not Yet Published.\\
   Updated 24th April, 2022.\\
   Page {\thepage} of \pageref{LastPage}.
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  %\includegraphics[width=48px,height=48px]{dave.jpeg}
  Measure your website readability!\\
  www.readability-score.com
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\begin{multicols*}{2}

\begin{tabularx}{8.4cm}{X}
\SetRowColor{DarkBackground}
\mymulticolumn{1}{x{8.4cm}}{\bf\textcolor{white}{Cell Counting by Hemocytometer}}  \tn
\SetRowColor{LightBackground}
\mymulticolumn{1}{p{8.4cm}}{\vspace{1px}\centerline{\includegraphics[width=5.1cm]{/web/www.cheatography.com/public/uploads/woozing_1650688229_Screenshot 2022-04-23 122636.png}}} \tn 
\hhline{>{\arrayrulecolor{DarkBackground}}-}
\end{tabularx}
\par\addvspace{1.3em}

\begin{tabularx}{8.4cm}{X}
\SetRowColor{DarkBackground}
\mymulticolumn{1}{x{8.4cm}}{\bf\textcolor{white}{Hemocytometer Counting Chamber}}  \tn
\SetRowColor{LightBackground}
\mymulticolumn{1}{p{8.4cm}}{\vspace{1px}\centerline{\includegraphics[width=5.1cm]{/web/www.cheatography.com/public/uploads/woozing_1650687957_Screenshot 2022-04-23 122537.png}}} \tn 
\hhline{>{\arrayrulecolor{DarkBackground}}-}
\end{tabularx}
\par\addvspace{1.3em}

\begin{tabularx}{8.4cm}{X}
\SetRowColor{DarkBackground}
\mymulticolumn{1}{x{8.4cm}}{\bf\textcolor{white}{Procedure}}  \tn
% Row 0
\SetRowColor{LightBackground}
\mymulticolumn{1}{x{8.4cm}}{-For an accurate cell count to be obtained, a uniform suspension containing single cells is necessary.} \tn 
% Row Count 3 (+ 3)
% Row 1
\SetRowColor{white}
\mymulticolumn{1}{x{8.4cm}}{{\bf{Obtain a uniform suspension of cells:}}} \tn 
% Row Count 4 (+ 1)
% Row 2
\SetRowColor{LightBackground}
\mymulticolumn{1}{x{8.4cm}}{1. Follow the typsinization/trypsin neutralization protocol for the specific cell type.} \tn 
% Row Count 6 (+ 2)
% Row 3
\SetRowColor{white}
\mymulticolumn{1}{x{8.4cm}}{2. Place the cell suspension in a suitably-sized conical centrifuge tube.} \tn 
% Row Count 8 (+ 2)
% Row 4
\SetRowColor{LightBackground}
\mymulticolumn{1}{x{8.4cm}}{3. a) Pipette the cell suspension up and down in the tube 5-7 times using a pipette with a small bore (5 ml or 10 ml pipette).} \tn 
% Row Count 11 (+ 3)
% Row 5
\SetRowColor{white}
\mymulticolumn{1}{x{8.4cm}}{3. b) For cells thawed from cryopreservation (in 1ml cryopreservation medium), pipette up and down 7-10 times using a one ml pipette.} \tn 
% Row Count 14 (+ 3)
% Row 6
\SetRowColor{LightBackground}
\mymulticolumn{1}{x{8.4cm}}{{\bf{Prepare a 1:1 dilution of the cell suspension in trypan blue: }}} \tn 
% Row Count 16 (+ 2)
% Row 7
\SetRowColor{white}
\mymulticolumn{1}{x{8.4cm}}{{\emph{Approximately 10 microliters of cell suspension will be required to charge one chamber of the hemocytometer.}}} \tn 
% Row Count 19 (+ 3)
% Row 8
\SetRowColor{LightBackground}
\mymulticolumn{1}{x{8.4cm}}{4. In a conical microfuge tube, add 10 microliters of 0.4\% trypan blue solution.} \tn 
% Row Count 21 (+ 2)
% Row 9
\SetRowColor{white}
\mymulticolumn{1}{x{8.4cm}}{5. Gently swirl (finger vortex) the cell suspension and remove 10 microliters of it using sterile technique.} \tn 
% Row Count 24 (+ 3)
% Row 10
\SetRowColor{LightBackground}
\mymulticolumn{1}{x{8.4cm}}{6. Combine the 10 microliters of cell suspension with the 10 microliters of trypan blue in the microfuge tube.} \tn 
% Row Count 27 (+ 3)
% Row 11
\SetRowColor{white}
\mymulticolumn{1}{x{8.4cm}}{7. Pipette up and down several times to ensure a uniform cell suspension using the same pipette tip and allow to stand for 5-15 minutes.} \tn 
% Row Count 30 (+ 3)
\end{tabularx}
\par\addvspace{1.3em}

\vfill
\columnbreak
\begin{tabularx}{8.4cm}{X}
\SetRowColor{DarkBackground}
\mymulticolumn{1}{x{8.4cm}}{\bf\textcolor{white}{Procedure (cont)}}  \tn
% Row 12
\SetRowColor{LightBackground}
\mymulticolumn{1}{x{8.4cm}}{{\bf{Load the haemocytometer: }}} \tn 
% Row Count 1 (+ 1)
% Row 13
\SetRowColor{white}
\mymulticolumn{1}{x{8.4cm}}{{\emph{The chamber should not be overfilled OR underfilled.}}} \tn 
% Row Count 3 (+ 2)
% Row 14
\SetRowColor{LightBackground}
\mymulticolumn{1}{x{8.4cm}}{8. Moisten and affix cover slip to the hemocytometer.} \tn 
% Row Count 5 (+ 2)
% Row 15
\SetRowColor{white}
\mymulticolumn{1}{x{8.4cm}}{9. Ensure the cover slip and hemocytometer are clean and grease-free (use alcohol to clean).} \tn 
% Row Count 7 (+ 2)
% Row 16
\SetRowColor{LightBackground}
\mymulticolumn{1}{x{8.4cm}}{10. A small amount of trypan blue-cell suspension is transferred  to one of the chambers of the hemocytometer by carefully touching  the cover slip at its edge  with the pipette tip and allowing each chamber to fill by capillary action.} \tn 
% Row Count 12 (+ 5)
% Row 17
\SetRowColor{white}
\mymulticolumn{1}{x{8.4cm}}{{\bf{Determine the number of cells (total and viable): }}} \tn 
% Row Count 14 (+ 2)
% Row 18
\SetRowColor{LightBackground}
\mymulticolumn{1}{x{8.4cm}}{11. View the cells under a microscope at 100x magnification.} \tn 
% Row Count 16 (+ 2)
% Row 19
\SetRowColor{white}
\mymulticolumn{1}{x{8.4cm}}{12. Under the microscope, you should see a grid of 9 squares.} \tn 
% Row Count 18 (+ 2)
% Row 20
\SetRowColor{LightBackground}
\mymulticolumn{1}{x{8.4cm}}{13. Focus the microscope on one of the 4 outer squares in the grid.} \tn 
% Row Count 20 (+ 2)
% Row 21
\SetRowColor{white}
\mymulticolumn{1}{x{8.4cm}}{14. The square should contain 16 smaller squares.} \tn 
% Row Count 21 (+ 1)
% Row 22
\SetRowColor{LightBackground}
\mymulticolumn{1}{x{8.4cm}}{15. Count all the cells in the four 1 mm corner squares.} \tn 
% Row Count 23 (+ 2)
% Row 23
\SetRowColor{white}
\mymulticolumn{1}{x{8.4cm}}{16. If there are too many or too few cells to count, repeat the procedure, either concentrating or diluting the original suspension as appropriate.} \tn 
% Row Count 26 (+ 3)
\hhline{>{\arrayrulecolor{DarkBackground}}-}
\end{tabularx}
\par\addvspace{1.3em}

\begin{tabularx}{8.4cm}{x{2.4 cm} x{5.6 cm} }
\SetRowColor{DarkBackground}
\mymulticolumn{2}{x{8.4cm}}{\bf\textcolor{white}{Types of Counting Method}}  \tn
% Row 0
\SetRowColor{LightBackground}
1. Logical count & -count the cells in the four corner squares and the middle square of the hemocytometer's grid \tn 
% Row Count 4 (+ 4)
% Row 1
\SetRowColor{white}
 & -can predict the cell count of the rest of the squares \tn 
% Row Count 6 (+ 2)
% Row 2
\SetRowColor{LightBackground}
 & -to make sure that the count is consistent and that cells aren't counted twice, you should only count cells on two of the square sides \tn 
% Row Count 11 (+ 5)
% Row 3
\SetRowColor{white}
 & -i.e. choose the cells on the left and top borders of the square while the ones on the right and top aren't included or the other way round \tn 
% Row Count 16 (+ 5)
% Row 4
\SetRowColor{LightBackground}
 & -be consistent throughout your counting procedure and to apply the same strategy every time, so you can compare your data \tn 
% Row Count 21 (+ 5)
% Row 5
\SetRowColor{white}
2. Absolute count & -count all the 9 squares in the hemocytometer \tn 
% Row Count 23 (+ 2)
% Row 6
\SetRowColor{LightBackground}
 & -ensures that you count everything and avoid error from the \seqsplit{generalization-assumption} mentioned above \tn 
% Row Count 27 (+ 4)
% Row 7
\SetRowColor{white}
 & -count the cells in the squares while following a zig-zag pattern \tn 
% Row Count 30 (+ 3)
\end{tabularx}
\par\addvspace{1.3em}

\vfill
\columnbreak
\begin{tabularx}{8.4cm}{x{2.4 cm} x{5.6 cm} }
\SetRowColor{DarkBackground}
\mymulticolumn{2}{x{8.4cm}}{\bf\textcolor{white}{Types of Counting Method (cont)}}  \tn
% Row 8
\SetRowColor{LightBackground}
 & -useful when there's a high cell concentration in the sample because of a pattern that's easy to follow \tn 
% Row Count 4 (+ 4)
% Row 9
\SetRowColor{white}
3. Quick count & -in a rush to do a cell count \tn 
% Row Count 6 (+ 2)
% Row 10
\SetRowColor{LightBackground}
 & -only count the cells in the top left square as well as the cells in the bottom right square \tn 
% Row Count 10 (+ 4)
% Row 11
\SetRowColor{white}
 & -won't get a result that is as representative as the other two methods mentioned above, but it can be a good way to spot-check your cell culture when in a hurry \tn 
% Row Count 16 (+ 6)
\hhline{>{\arrayrulecolor{DarkBackground}}--}
\end{tabularx}
\par\addvspace{1.3em}

\begin{tabularx}{8.4cm}{X}
\SetRowColor{DarkBackground}
\mymulticolumn{1}{x{8.4cm}}{\bf\textcolor{white}{The Logical Count}}  \tn
\SetRowColor{LightBackground}
\mymulticolumn{1}{p{8.4cm}}{\vspace{1px}\centerline{\includegraphics[width=5.1cm]{/web/www.cheatography.com/public/uploads/woozing_1650798914_Screenshot 2022-04-24 190036.png}}} \tn 
\hhline{>{\arrayrulecolor{DarkBackground}}-}
\end{tabularx}
\par\addvspace{1.3em}

\begin{tabularx}{8.4cm}{X}
\SetRowColor{DarkBackground}
\mymulticolumn{1}{x{8.4cm}}{\bf\textcolor{white}{The Absolute Count}}  \tn
\SetRowColor{LightBackground}
\mymulticolumn{1}{p{8.4cm}}{\vspace{1px}\centerline{\includegraphics[width=5.1cm]{/web/www.cheatography.com/public/uploads/woozing_1650798934_Screenshot 2022-04-24 191452.png}}} \tn 
\hhline{>{\arrayrulecolor{DarkBackground}}-}
\end{tabularx}
\par\addvspace{1.3em}

\begin{tabularx}{8.4cm}{X}
\SetRowColor{DarkBackground}
\mymulticolumn{1}{x{8.4cm}}{\bf\textcolor{white}{The Quick Count}}  \tn
\SetRowColor{LightBackground}
\mymulticolumn{1}{p{8.4cm}}{\vspace{1px}\centerline{\includegraphics[width=5.1cm]{/web/www.cheatography.com/public/uploads/woozing_1650798954_Screenshot 2022-04-24 191423.png}}} \tn 
\hhline{>{\arrayrulecolor{DarkBackground}}-}
\end{tabularx}
\par\addvspace{1.3em}

\begin{tabularx}{8.4cm}{X}
\SetRowColor{DarkBackground}
\mymulticolumn{1}{x{8.4cm}}{\bf\textcolor{white}{Cell Counting Accuracy}}  \tn
% Row 0
\SetRowColor{LightBackground}
\mymulticolumn{1}{x{8.4cm}}{{\bf{The total number of cells overlying one 1 mm2 should be between 15 and 50}}} \tn 
% Row Count 2 (+ 2)
% Row 1
\SetRowColor{white}
\mymulticolumn{1}{x{8.4cm}}{-If the number of cells per 1 mm2 exceeds 50, dilute the sample and count again} \tn 
% Row Count 4 (+ 2)
% Row 2
\SetRowColor{LightBackground}
\mymulticolumn{1}{x{8.4cm}}{-If the number of cells per 1 mm2 is less than 15, use a less diluted sample} \tn 
% Row Count 6 (+ 2)
% Row 3
\SetRowColor{white}
\mymulticolumn{1}{x{8.4cm}}{-If less dilute samples are not available, count cells on both sides of the hemocytometer (8 x 1 mm2 areas)} \tn 
% Row Count 9 (+ 3)
% Row 4
\SetRowColor{LightBackground}
\mymulticolumn{1}{x{8.4cm}}{} \tn 
% Row Count 9 (+ 0)
% Row 5
\SetRowColor{white}
\mymulticolumn{1}{x{8.4cm}}{{\bf{Keep a separate count of viable and non-viable cells}}} \tn 
% Row Count 11 (+ 2)
% Row 6
\SetRowColor{LightBackground}
\mymulticolumn{1}{x{8.4cm}}{-If \textgreater{} 25\% of cells are non-viable, the culture is not being maintained on the appropriate amount of media. } \tn 
% Row Count 14 (+ 3)
% Row 7
\SetRowColor{white}
\mymulticolumn{1}{x{8.4cm}}{-Reincubate the culture and adjust the volume of media according to the confluency of the cells and the appearance of the media.} \tn 
% Row Count 17 (+ 3)
% Row 8
\SetRowColor{LightBackground}
\mymulticolumn{1}{x{8.4cm}}{-Include cells on top and left touching middle line.} \tn 
% Row Count 19 (+ 2)
% Row 9
\SetRowColor{white}
\mymulticolumn{1}{x{8.4cm}}{-The cells touching middle line at bottom and right are not counted.} \tn 
% Row Count 21 (+ 2)
\hhline{>{\arrayrulecolor{DarkBackground}}-}
\end{tabularx}
\par\addvspace{1.3em}

\begin{tabularx}{8.4cm}{X}
\SetRowColor{DarkBackground}
\mymulticolumn{1}{x{8.4cm}}{\bf\textcolor{white}{Loading Cell Suspension Mixture}}  \tn
\SetRowColor{LightBackground}
\mymulticolumn{1}{p{8.4cm}}{\vspace{1px}\centerline{\includegraphics[width=5.1cm]{/web/www.cheatography.com/public/uploads/woozing_1650799108_Screenshot 2022-04-24 191745.png}}} \tn 
\hhline{>{\arrayrulecolor{DarkBackground}}-}
\end{tabularx}
\par\addvspace{1.3em}

\begin{tabularx}{8.4cm}{X}
\SetRowColor{DarkBackground}
\mymulticolumn{1}{x{8.4cm}}{\bf\textcolor{white}{Cell Density}}  \tn
\SetRowColor{LightBackground}
\mymulticolumn{1}{p{8.4cm}}{\vspace{1px}\centerline{\includegraphics[width=5.1cm]{/web/www.cheatography.com/public/uploads/woozing_1650799161_Screenshot 2022-04-24 191846.png}}} \tn 
\hhline{>{\arrayrulecolor{DarkBackground}}-}
\end{tabularx}
\par\addvspace{1.3em}

\begin{tabularx}{8.4cm}{X}
\SetRowColor{DarkBackground}
\mymulticolumn{1}{x{8.4cm}}{\bf\textcolor{white}{Trypan Blue}}  \tn
% Row 0
\SetRowColor{LightBackground}
\mymulticolumn{1}{x{8.4cm}}{-"vital stain", excluded dead cells from living cells} \tn 
% Row Count 2 (+ 2)
% Row 1
\SetRowColor{white}
\mymulticolumn{1}{x{8.4cm}}{-living cells appear colourless and bright (refractile) under phase contrast.} \tn 
% Row Count 4 (+ 2)
% Row 2
\SetRowColor{LightBackground}
\mymulticolumn{1}{x{8.4cm}}{-\textgreater{}charged trypan blue molecules do not penetrate into viable cells because of the integrity of the membrane} \tn 
% Row Count 7 (+ 3)
% Row 3
\SetRowColor{white}
\mymulticolumn{1}{x{8.4cm}}{-dead cells stain blue and are non-refractile} \tn 
% Row Count 8 (+ 1)
% Row 4
\SetRowColor{LightBackground}
\mymulticolumn{1}{x{8.4cm}}{-\textgreater{}non-viable cells, allow the free entry of trypan blue, thereby staining the cell interior blue} \tn 
% Row Count 10 (+ 2)
\hhline{>{\arrayrulecolor{DarkBackground}}-}
\end{tabularx}
\par\addvspace{1.3em}

\begin{tabularx}{8.4cm}{X}
\SetRowColor{DarkBackground}
\mymulticolumn{1}{x{8.4cm}}{\bf\textcolor{white}{Living Cells and Dead Cells under the Microscope}}  \tn
\SetRowColor{LightBackground}
\mymulticolumn{1}{p{8.4cm}}{\vspace{1px}\centerline{\includegraphics[width=5.1cm]{/web/www.cheatography.com/public/uploads/woozing_1650799409_Screenshot 2022-04-24 192205.png}}} \tn 
\hhline{>{\arrayrulecolor{DarkBackground}}-}
\end{tabularx}
\par\addvspace{1.3em}

\begin{tabularx}{8.4cm}{X}
\SetRowColor{DarkBackground}
\mymulticolumn{1}{x{8.4cm}}{\bf\textcolor{white}{Counting Cells}}  \tn
\SetRowColor{LightBackground}
\mymulticolumn{1}{p{8.4cm}}{\vspace{1px}\centerline{\includegraphics[width=5.1cm]{/web/www.cheatography.com/public/uploads/woozing_1650799494_Screenshot 2022-04-24 192431.png}}} \tn 
\hhline{>{\arrayrulecolor{DarkBackground}}-}
\end{tabularx}
\par\addvspace{1.3em}

\begin{tabularx}{8.4cm}{X}
\SetRowColor{DarkBackground}
\mymulticolumn{1}{x{8.4cm}}{\bf\textcolor{white}{Corner Squares}}  \tn
% Row 0
\SetRowColor{LightBackground}
\mymulticolumn{1}{x{8.4cm}}{-each of the 16 smaller squares will be 1 mm/4 = 0.25 mm in width and 0.25 mm x 0.25 mm = 0.0625 mm2 in area (or 1 mm2/16 = 0.0625 mm2)} \tn 
% Row Count 3 (+ 3)
% Row 1
\SetRowColor{white}
\mymulticolumn{1}{x{8.4cm}}{-cells that are 10 μm or more should be counted in these corner squares} \tn 
% Row Count 5 (+ 2)
% Row 2
\SetRowColor{LightBackground}
\mymulticolumn{1}{x{8.4cm}}{-i.e. white blood cells} \tn 
% Row Count 6 (+ 1)
\hhline{>{\arrayrulecolor{DarkBackground}}-}
\end{tabularx}
\par\addvspace{1.3em}

\begin{tabularx}{8.4cm}{X}
\SetRowColor{DarkBackground}
\mymulticolumn{1}{x{8.4cm}}{\bf\textcolor{white}{When to Use Corner Squares}}  \tn
\SetRowColor{LightBackground}
\mymulticolumn{1}{p{8.4cm}}{\vspace{1px}\centerline{\includegraphics[width=5.1cm]{/web/www.cheatography.com/public/uploads/woozing_1650799698_Screenshot 2022-04-24 192747.png}}} \tn 
\hhline{>{\arrayrulecolor{DarkBackground}}-}
\end{tabularx}
\par\addvspace{1.3em}

\begin{tabularx}{8.4cm}{X}
\SetRowColor{DarkBackground}
\mymulticolumn{1}{x{8.4cm}}{\bf\textcolor{white}{Central Square}}  \tn
% Row 0
\SetRowColor{LightBackground}
\mymulticolumn{1}{x{8.4cm}}{-each of the 25 smaller squares will be 1 mm/5 = 0.2 mm in width and 0.2 mm x 0.2 mm = 0.04 mm2 in area (or 1 mm 2/25 = 0.04 mm2)} \tn 
% Row Count 3 (+ 3)
% Row 1
\SetRowColor{white}
\mymulticolumn{1}{x{8.4cm}}{-each of the 25 smaller squares contains 16 even smaller squares which measure: 0.2 mm/4 = 0.05 mm in width and 0.05 mm x 0.05 mm = 0.0025 mm2 = 2500 μm2 (or 0.04 mm2/16 = 0.0025 mm2)} \tn 
% Row Count 7 (+ 4)
% Row 2
\SetRowColor{LightBackground}
\mymulticolumn{1}{x{8.4cm}}{-Cells that are 10 μm or smaller should be counted in the central square – sometimes even in one of the smaller squares inside the central square} \tn 
% Row Count 10 (+ 3)
% Row 3
\SetRowColor{white}
\mymulticolumn{1}{x{8.4cm}}{-i.e. red blood cells, platelets, most types of yeast, and sperm cells} \tn 
% Row Count 12 (+ 2)
\hhline{>{\arrayrulecolor{DarkBackground}}-}
\end{tabularx}
\par\addvspace{1.3em}

\begin{tabularx}{8.4cm}{X}
\SetRowColor{DarkBackground}
\mymulticolumn{1}{x{8.4cm}}{\bf\textcolor{white}{When to Use Central Squares}}  \tn
\SetRowColor{LightBackground}
\mymulticolumn{1}{p{8.4cm}}{\vspace{1px}\centerline{\includegraphics[width=5.1cm]{/web/www.cheatography.com/public/uploads/woozing_1650800401_Screenshot 2022-04-24 193926.png}}} \tn 
\hhline{>{\arrayrulecolor{DarkBackground}}-}
\end{tabularx}
\par\addvspace{1.3em}

\begin{tabularx}{8.4cm}{X}
\SetRowColor{DarkBackground}
\mymulticolumn{1}{x{8.4cm}}{\bf\textcolor{white}{Calculation}}  \tn
% Row 0
\SetRowColor{LightBackground}
\mymulticolumn{1}{x{8.4cm}}{-count 4 corner squares and calculate the average} \tn 
% Row Count 1 (+ 1)
% Row 1
\SetRowColor{white}
\mymulticolumn{1}{x{8.4cm}}{-each large square of the haemocytometer, with cover slip in place, represents a total volume of 0.1 mm3 (1.0mm X1.0mmX 0.1mm) or 10-4 cm3} \tn 
% Row Count 4 (+ 3)
% Row 2
\SetRowColor{LightBackground}
\mymulticolumn{1}{x{8.4cm}}{-1 cm3 = 1 mL} \tn 
% Row Count 5 (+ 1)
% Row 3
\SetRowColor{white}
\mymulticolumn{1}{x{8.4cm}}{} \tn 
% Row Count 5 (+ 0)
% Row 4
\SetRowColor{LightBackground}
\mymulticolumn{1}{x{8.4cm}}{{\bf{\% Cell Viability:}}} \tn 
% Row Count 6 (+ 1)
% Row 5
\SetRowColor{white}
\mymulticolumn{1}{x{8.4cm}}{= {[}Total Viable cells (Unstained) / Total cells (Viable + Dead){]}  X 100\%} \tn 
% Row Count 8 (+ 2)
% Row 6
\SetRowColor{LightBackground}
\mymulticolumn{1}{x{8.4cm}}{{\bf{Viable Cells/ml:}}} \tn 
% Row Count 9 (+ 1)
% Row 7
\SetRowColor{white}
\mymulticolumn{1}{x{8.4cm}}{= Average viable cell count per square x Dilution Factor x 10\textasciicircum{}4} \tn 
% Row Count 11 (+ 2)
% Row 8
\SetRowColor{LightBackground}
\mymulticolumn{1}{x{8.4cm}}{{\bf{Average viable cell count per square: }}} \tn 
% Row Count 12 (+ 1)
% Row 9
\SetRowColor{white}
\mymulticolumn{1}{x{8.4cm}}{= Total number of viable cells from the number of squares / number of squares} \tn 
% Row Count 14 (+ 2)
% Row 10
\SetRowColor{LightBackground}
\mymulticolumn{1}{x{8.4cm}}{{\bf{Dilution Factor:}}} \tn 
% Row Count 15 (+ 1)
% Row 11
\SetRowColor{white}
\mymulticolumn{1}{x{8.4cm}}{= Total Volume (Volume of sample + Volume of diluting liquid) / Volume of sample.} \tn 
% Row Count 17 (+ 2)
% Row 12
\SetRowColor{LightBackground}
\mymulticolumn{1}{x{8.4cm}}{{\bf{Total viable cells/Sample: }}} \tn 
% Row Count 18 (+ 1)
% Row 13
\SetRowColor{white}
\mymulticolumn{1}{x{8.4cm}}{= Viable Cells/ml x The original volume of fluid from which the cell sample was removed.} \tn 
% Row Count 20 (+ 2)
% Row 14
\SetRowColor{LightBackground}
\mymulticolumn{1}{x{8.4cm}}{{\bf{Volume of media needed:}}} \tn 
% Row Count 21 (+ 1)
% Row 15
\SetRowColor{white}
\mymulticolumn{1}{x{8.4cm}}{= (Number of cells needed/Total number of viable cells) x 10\textasciicircum{}4} \tn 
% Row Count 23 (+ 2)
% Row 16
\SetRowColor{LightBackground}
\mymulticolumn{1}{x{8.4cm}}{{\bf{Cell density (cell/mL):}}} \tn 
% Row Count 24 (+ 1)
% Row 17
\SetRowColor{white}
\mymulticolumn{1}{x{8.4cm}}{= average cell count per square x 10\textasciicircum{}4 x dilution factor} \tn 
% Row Count 26 (+ 2)
\hhline{>{\arrayrulecolor{DarkBackground}}-}
\SetRowColor{LightBackground}
\mymulticolumn{1}{x{8.4cm}}{How to count: \seqsplit{https://www.youtube.com/watch?v=pP0xERLUhyc}}  \tn 
\hhline{>{\arrayrulecolor{DarkBackground}}-}
\end{tabularx}
\par\addvspace{1.3em}


% That's all folks
\end{multicols*}

\end{document}