ADD a bunch of figures

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Nathan Dwarshuis 2021-07-23 17:14:45 -04:00
parent 14d598b1b7
commit 7cd0ca05ba
1 changed files with 113 additions and 8 deletions

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@ -61,6 +61,7 @@
\newacronym{bca}{BCA}{bicinchoninic acid assay}
\newacronym{bsa}{BSA}{bovine serum albumin}
\newacronym{stp}{STP}{streptavidin}
\newacronym{stppe}{STP-PE}{streptavidin-phycoerythrin}
\newacronym{snb}{SNB}{sulfo-nhs-biotin}
\newacronym{cug}{CuG}{Cultispher G}
\newacronym{cus}{CuS}{Cultispher S}
@ -132,6 +133,8 @@
\newcommand{\inlinecode}{\texttt}
\newcommand{\subcap}[2]{\subref{#1}) #2}
%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%
% ditto for environments
@ -662,6 +665,16 @@ better retention of memory phenotype compared to current bead-based methods.
\section{methods}
\subsection{dms functionalization}
\begin{figure*}[ht!]
\begingroup
\includegraphics{../figures/dms_flowchart.png}
\endgroup
\caption[\gls{dms} Flowchart]{Overview of \gls{dms} manufacturing process.}
\label{fig:dms_flowchart}
\end{figure*}
Gelatin microcarriers (\gls{cus} or \gls{cug}, GE Healthcare, DG-2001-OO and
DG-0001-OO) were suspended at \SI{20}{\mg\per\ml} in 1X \gls{pbs} and
autoclaved. All subsequent steps were done aseptically, and all reactions were
@ -781,7 +794,7 @@ threshold. Media glucose was measured using a \product{GlucCell glucose
% added to a final concentration of \SI{50}{\ug\per\ml} during media addition.
Cells on the \glspl{dms} were visualized by adding \SI{0.5}{\ul}
\product{\gls{stp}-\gls{pe}}{\bl}{405204} and \SI{2}{ul}
\product{\gls{stppe}}{\bl}{405204} and \SI{2}{ul}
\product{\acd{45}-\gls{af647}}{\bl}{368538}, incubating for \SI{1}{\hour}, and
imaging on a spinning disk confocal microscope.
@ -868,21 +881,113 @@ context of pure error). Statistical significance was evaluated at $\upalpha$ =
\subsection{DMSs can be fabricated in a controlled manner}
\begin{figure*}[ht!]
\begingroup
\includegraphics{../figures/dms_coating.png}
\phantomsubcaption\label{fig:stp_carrier_fitc}
\phantomsubcaption\label{fig:mab_carrier_fitc}
\phantomsubcaption\label{fig:cug_vs_cus}
\phantomsubcaption\label{fig:biotin_coating}
\phantomsubcaption\label{fig:stp_coating}
\phantomsubcaption\label{fig:mab_coating}
\endgroup
\caption[\gls{dms} Coating]
{\gls{dms} functionalization results.
\subcap{fig:stp_carrier_fitc}{\gls{stp}-coated or uncoated \glspl{dms}
treated with biotin-FITC and imaged using a lightsheet microscope.}
\subcap{fig:mab_carrier_fitc}{\gls{mab}-coated or \gls{stp}-coated
\glspl{dms} treated with \anti{\gls{igg}} \glspl{mab} and imaged using a
lightsheet microscope.} \subcap{fig:cug_vs_cus}{Bound \gls{stp} surface
density on either \gls{cus} or \gls{cug} microcarriers. Surface density
was estimated using the properties in~\cref{tab:carrier_props}} Total
binding curve of \subcap{fig:biotin_coating}{biotin},
\subcap{fig:stp_coating}{\gls{stp}}, and
\subcap{fig:mab_coating}{\glspl{mab}} as a function of biotin added. }
\label{fig:dms_flowchart}
\end{figure*}
% TODO these caption titles suck
% TODO combine this DOE figure into one interaction plot
\begin{table}[!h] \centering
\caption{Properties of the microcarriers used}
\label{tab:ci_treat}
\label{tab:carrier_props}
\input{../tables/carrier_properties.tex}
\end{table}
% fabrication flow chart
% microcarrier properties
% green carrier figure
% quality control figure
% DMS manufacturing timing figure
\begin{figure*}[ht!]
\begingroup
\includegraphics{../figures/dms_qc.png}
\phantomsubcaption\label{fig:dms_qc_doe}
\phantomsubcaption\label{fig:dms_qc_ph}
\phantomsubcaption\label{fig:dms_qc_washes}
\phantomsubcaption\label{fig:dms_snb_decay_curves}
\endgroup
\caption[\gls{dms} Quality Control]
{\gls{dms} quality control investigation and development
\subcap{fig:dms_qc_doe}{\gls{doe} investigating the effect of initial mass
of microcarriers, reaction temperature, and biotin concentration on
biotin attachment.}
\subcap{fig:dms_qc_ph}{Effect of reaction ph on biotin attachment.}
\subcap{fig:dms_qc_washes}{effect of post-autoclave washing of the
microcarriers on biotin attachment.}
\subcap{fig:dms_snb_decay_curves}{Hydrolysis curves of \gls{snb} in
\gls{pbs} of differing pH.}
All statistical tests where p-values are noted are given by two-tailed t
tests.
}
\label{fig:dms_flowchart}
\end{figure*}
\begin{table}[!h] \centering
\caption{Properties of the microcarriers used}
\label{tab:carrier_props}
\input{../tables/carrier_properties.tex}
\end{table}
\begin{figure*}[ht!]
\begingroup
\includegraphics{../figures/dms_timing.png}
\phantomsubcaption\label{fig:dms_biotin_rxn_mass}
\phantomsubcaption\label{fig:dms_biotin_rxn_frac}
\phantomsubcaption\label{fig:dms_stp_per_time}
\endgroup
\caption[\gls{dms} Reaction timing]
{Reaction kinetics for microcarrier functionalization.
\subcap{fig:dms_biotin_rxn_mass}{Biotin mass bound per time}
\subcap{fig:dms_biotin_rxn_frac}{Fraction of input biotin bound per time}
\subcap{fig:dms_stp_per_time}{\Gls{stp} bound per time.}
}
\label{fig:dms_flowchart}
\end{figure*}
\subsection{DMSs can efficiently expand T cells compared to beads}
% cells growing on DMSs
% TODO make sure the day on these is correct
\begin{figure*}[ht!]
\begingroup
\includegraphics{../figures/cells_on_dms.png}
\phantomsubcaption\label{fig:dms_cells_phase}
\phantomsubcaption\label{fig:dms_cells_fluor}
\endgroup
\caption[\gls{dms} Reaction timing]
{Cells grow in tight clusters in and around functionalized \gls{dms}.
\subcap{fig:dms_cells_phase}{Phase-contrast image of T cells growing on
\glspl{dms} on day 7}
\subcap{fig:dms_cells_fluor}{Confocal images of T cells in varying z-planes
growing on \glspl{dms} on day 9. \Glspl{dms} were stained using
\gls{stppe} (red) and T cells were stained using \acd{45}-\gls{af647}.}
}
\label{fig:dms_flowchart}
\end{figure*}
% 3-donor expansion figure
\subsection{DMSs lead to greater expansion and memory and CD4+ phenotypes}