ADD interior quantification section

tex/thesis.tex

@@ -803,7 +803,7 @@ adding \gls{fitcbt} to the \gls{stp}-coated \glspl{dms}, resuspending in
 staining with \product{\anti{\gls{igg}}-\gls{fitc}}{\bl}{406001}, incubating for
 \SI{30}{\minute}, washing with \gls{pbs}, and imaging on a confocal microscope.
 
-\subsection{t cell culture}
+\subsection{t cell culture}\label{sec:tcellculture}
 
 % TODO verify countess product number
 Cryopreserved primary human T cells were either obtained as sorted
@@ -838,7 +838,24 @@ Cells on the \glspl{dms} were visualized by adding \SI{0.5}{\ul}
 \product{\acd{45}-\gls{af647}}{\bl}{368538}, incubating for \SI{1}{\hour}, and
 imaging on a spinning disk confocal microscope.
 
-% METHOD mtt staining and interior quantification
+\subsection{Quantifying cells on DMS interior}
+
+% TODO add a product number to MTT (if I can find it)
+Cells were stained and visualized using \gls{mtt}. \glspl{dms} with attached and
+loosely attached cells were sampled as desired and filtered through a
+\SI{40}{\um} cell strainer. While still in the cell strainer, \glspl{dms} were
+washed twice with \gls{pbs} and then dried by pulling liquid through the bottom
+of the cell strainer via a micropipette and dabbing with a KimWipe. \glspl{dms}
+were transferred to a 24 well plate with \SI{400}{\ul} media. \SI{40}{\ul}
+\gls{mtt} was added to each well and allowed to incubate for \SI{3}{\hour},
+after which \glspl{dms} with cell were visualized via a brightfield microscope.
+
+To quantify cells on the interior of \glspl{dms}, cells and \glspl{dms} were
+isolated analogously to those for the \gls{mtt} stain up until the drying step.
+Cells were then transferred to a tube containing \SI{400}{\ul} at
+\SI{5}{\mg\per\ml} dispase solution. \glspl{dms} were incubated and rotated for
+\SI{45}{\minute} at \SI{37}{\degreeCelsius}, after which cells were counted as
+already described in \cref{sec:tcellculture}.
 
 \subsection{quantification of apoptosis using Annexin-V}