%!TEX root=../labNotes.tex

\subsection*{09 September 2014}

\begin{enumerate}
	\item MA104 cell split --- Passage 60, from Passage 59, Flask A
		\begin{enumerate}
			\item Aspirated cell culture medium
			\item Rinsed cells in $10$mL 1x PBS; aspirated PBS
			\item Rinsed cells in $2$mL $0.05$\% trypsin; aspirated trypsin
			\item Bathed cells in $2$mL $0.05$\% trypsin
			\item Incubated cells at $37^{\circ}$C until all cells detached from flask
			\item Added $18$mL complete M199 to flask
			\item Added to $3$ T150 flasks $15$mL complete M199/$10$mL cell mix; $20$mL complete M199/$5$mL cell mix; and $20$mL complete M199/$5$mL cell mix. Respectively, flasks A, B, and C
			\item Gently shook flasks to distribute cells evenly
			\item Incubated at $37^{\circ}$C
		\end{enumerate}
\end{enumerate}

\subsection*{12 September 2014}

\begin{enumerate}
	\item Plated MA104 cells --- Passage 60, Flask A
		\begin{enumerate}
			\item Aspirated cell culture medium
			\item Rinsed cells in $10$mL 1x PBS; aspirated PBS
			\item Rinsed cells in $5$mL $0.05$\% trypsin; aspirated trypsin
			\item Bathed cells in $5$mL $0.05$\% trypsin
			\item Incubated cells at $37^{\circ}$C until all cells detached from flask
			\item Added $15$mL complete M199 to flask
			\item Took cell count by combining $10\mu$L cell mixture with $10\mu$L trypan blue:
			
				\begin{align*}
				\text{[cells]} &= \frac{3.91\e{5}\text{ cells}}{1\text{mL}} \\
				\frac{\text{cells}}{\text{flask}} &= \frac{3.91\e{5}\text{ cells}}{1\text{mL}} \cdot 20\text{mL} &= \frac{7.62\e{6}\text{ cells}}{20\text{mL}}\\
				\frac{\text{cells}}{10\text{mL cell mix}} &= \frac{7.62\e{6}\text{ cells}}{20\text{mL}}\cdot \frac{1}{2} &= \frac{3.81\e{6}\text{ cells}}{10\text{mL}}\\
				\frac{\text{cells}}{75\text{mL vial}} &= \frac{3.81\e{6}\text{ cells}}{75\text{mL}} &= \frac{5.08\e{4}\text{ cells}}{\text{mL}}\\
				\frac{\text{cells}}{3\text{mL well}} &= \frac{5.08\e{4}\text{ cells}}{\text{mL}} \cdot 3\text{mL} &= \frac{1.52\e{5}\text{ cells}}{\text{well}}\\
				\end{align*}
			\item Added $65$mL complete M199 and $10$mL cell mixture to $125$mL conical vial for final volume of $75$mL
			\item Transferred $3$mL solution to each well of 4 6-well plates
			\item Spread cells evenly by shaking
			\item Incubated at $37^{\circ}$C
		\end{enumerate}
\end{enumerate}