From 02112996878a256a7f0c2cc80a264ba4fe91678d Mon Sep 17 00:00:00 2001 From: "lorenzo.zolfanelli93" Date: Fri, 28 Aug 2020 05:04:14 +0000 Subject: [PATCH] Update on Overleaf. --- chapters/3-methods.tex | 21 ++++++++++++++++----- images/alexa647.png | Bin 0 -> 122683 bytes 2 files changed, 16 insertions(+), 5 deletions(-) create mode 100644 images/alexa647.png diff --git a/chapters/3-methods.tex b/chapters/3-methods.tex index da4c035..5bc85a4 100644 --- a/chapters/3-methods.tex +++ b/chapters/3-methods.tex @@ -609,10 +609,10 @@ regione di moltiplicazione, dove, grazie a un'elevata differenza di potenziale, viene massimizzata la possibilità di generare nuovi elettroni per ionizzazione da impatto. Il segnale (sotto forma di cariche accumulate) che a questo punto viene trasferito allo stadio di integrazione e lettura -del CCD, presentà già un elevato guadagno senza dover far ricorso a +del CCD, presenta già un elevato guadagno senza dover far ricorso a ulteriori stadi di amplificazione elettronica. Per operare correttamente il sensore EMCCD deve essere raffreddato -a temperature $< \SI{-60}{\Celsius}$, in modo da ridurre il più +a temperature $< \SI{-60}{\celsius}$, in modo da ridurre il più possibile gli elettroni prodotti termicamente (corrente di buio), che possono subire lo stesso processo di moltiplicazione dei fotoelettroni. @@ -624,8 +624,8 @@ Quindi la presenza di fluorofori liberi in soluzione provoca un aumento consistente del rumore dovuto alla fluorescenza fuori fuoco. I fluorofori che si intende utilizzare per lo studio della -meccanotrasduzione appartegono alla famiglia Alexa Fluor (Molecular Probes). -Si tratta di molecole organiche sintentica particolarmente fotostabili +meccanotrasduzione appartengono alla famiglia Alexa Fluor (Molecular Probes). +Si tratta di molecole organiche sintetica particolarmente fotostabili e con resa quantica elevata. In commercio si trovano versioni funzionalizzate legate a svariate molecole e kit di coniugazione che consentono di legare semplicemente e rapidamente @@ -634,7 +634,7 @@ Tenendo conto delle caratteristiche dell'apparato è possibile utilizzare i fluorofori Alexa Fluor 647, 532 e 488. In figura \ref{fig:alexa_spectra} si riportano gli spettri di eccitazione e emissione. -\begin{figure} +\begin{figure}[ht] \centering \includegraphics[width=0.8\linewidth]{images/alexa.png} \caption{Spettri di eccitazione (linea tratteggiata) ed emissione @@ -643,8 +643,19 @@ si riportano gli spettri di eccitazione e emissione. \label{fig:alexa_spectra} \end{figure} +Per testare la capacità di visualizzare singoli fluorofori e stimare le +prestazioni del sistema utilizziamo dei campioni in cui una distribuzione +di Alexa 647 è immobilizzata sul vetrino coprioggetti (vedi appendice +\ref{app:protocols}, protocollo \ref{proto:alexa}). +\begin{figure}[ht] + \centering + \includegraphics{images/alexa647.png} + \caption{Caption} + \label{fig:my_label} +\end{figure} + \section{TIRF e illuminazione a modi di galleria} \label{sec:gallery_mode} diff --git a/images/alexa647.png b/images/alexa647.png new file mode 100644 index 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