From 3e5cceb066263d662fd5c9dd877b06e444fb8c09 Mon Sep 17 00:00:00 2001 From: "lorenzo.zolfanelli93" Date: Mon, 15 Jun 2020 16:15:47 +0000 Subject: [PATCH] Update on Overleaf. --- .travis.yml | 22 ++ README.md | 3 + chapters/1-introduction.tex | 111 ++++++++ chapters/2-methods.tex | 152 +++++++++++ deploy_key.enc | Bin 0 -> 1680 bytes images/Setup.pdf | Bin 0 -> 162820 bytes images/unifi.pdf | Bin 0 -> 48446 bytes images_src/unifi.svg | 520 ++++++++++++++++++++++++++++++++++++ main.tex | 56 ++++ references.bib | 16 ++ setup-fig/fig1.tex | 8 + texlive/texlive.profile | 10 + texlive/texlive_install.sh | 33 +++ texlive/texlive_packages | 21 ++ titlepage.tex | 74 +++++ 15 files changed, 1026 insertions(+) create mode 100644 .travis.yml create mode 100644 README.md create mode 100644 chapters/1-introduction.tex create mode 100644 chapters/2-methods.tex create mode 100644 deploy_key.enc create mode 100644 images/Setup.pdf create mode 100644 images/unifi.pdf create mode 100644 images_src/unifi.svg create mode 100644 main.tex create mode 100644 references.bib create mode 100644 setup-fig/fig1.tex create mode 100644 texlive/texlive.profile create mode 100644 texlive/texlive_install.sh create mode 100644 texlive/texlive_packages create mode 100644 titlepage.tex diff --git a/.travis.yml b/.travis.yml new file mode 100644 index 0000000..50112e2 --- /dev/null +++ b/.travis.yml @@ -0,0 +1,22 @@ +language: generic +dist: bionic +install: +- source ./texlive/texlive_install.sh +cache: + directories: + - "/tmp/texlive" + - "$HOME/.texlive" +before_install: +- openssl aes-256-cbc -K $encrypted_baf79bfacc4c_key -iv $encrypted_baf79bfacc4c_iv + -in deploy_key.enc -out deploy_key -d +- chmod 600 deploy_key +before_script: cd $TRAVIS_BUILD_DIR +script: +- texliveonfly main.tex +- latexmk -pdf main.tex +after_script: +- export PATH=/tmp/texlive/bin/x86_64-linux:$PATH +- tlmgr list --only-installed | grep -oP 'i \K.+?(?=:)' +after_success: +- rsync -e "ssh -p ${SFTP_PORT} -o StrictHostKeyChecking=no -i./deploy_key" main.pdf ${SFTP_HOST}:${SFTP_PREFIX}_${TRAVIS_BRANCH}_latest.pdf +- rsync -e "ssh -p ${SFTP_PORT} -o StrictHostKeyChecking=no -i./deploy_key" main.pdf ${SFTP_HOST}:${SFTP_PREFIX}_${TRAVIS_BRANCH}_${TRAVIS_JOB_NUMBER}.pdf diff --git a/README.md b/README.md new file mode 100644 index 0000000..15d3bf1 --- /dev/null +++ b/README.md @@ -0,0 +1,3 @@ +### Lorenzo Zolfanelli's Master Thesis + +[![Build Status](https://api.travis-ci.com/zolfariot/Master-Thesis.svg?token=RsJykoGjZjDowDKAdAzt&branch=master)](https://travis-ci.com/zolfariot/Master-Thesis) [Last Build](http://posta.zolfanelli.net/Tesi/LZ_Thesis_master_latest.pdf) diff --git a/chapters/1-introduction.tex b/chapters/1-introduction.tex new file mode 100644 index 0000000..5840a69 --- /dev/null +++ b/chapters/1-introduction.tex @@ -0,0 +1,111 @@ +\chapter{Introduzione} + + +Gli stimoli meccanici rivestono nell'ambito dei sistemi biologici un +ruolo importante nel determinare il corretto funzionamento di cellule, +tessuti e organismi complessi. + +Mentre tradizionalmente la Biologia si è occupata di +studiare come processi cellulari e inter-cellulari fossero regolati +dallo scambio di molecole biologiche, il ruolo degli stimoli +meccanici è stato a lungo ritenuto marginale nella descrizione di +questi processi. + +Lo sviluppo di tecniche sempre più avanzate e precise per la +visualizzazione e la manipolazione di molecole all'interno di campioni +biologici ha iniziato a mutare questa concezione: oggi possiamo +indagare nel dettaglio il funzionamento dei motori +molecolari all'interno delle nostre cellule o +misurare come variazioni nella tensione applicata ad un polimero +possano indurre una riorganizzazione strutturale nello stesso e +cambiarne le proprietà biochimiche. + +Per molti processi biologici il ruolo della forza è fondamentale, +ad esempio nei complessi proteici che legano tra di loro le cellule +in un tessuto. Questi si comportano come complesse macchine in grado +di elaborare stimoli di tipo biochimico e meccanico, comunicando e +interferendo con il resto delle funzioni cellulari. + +Le pinzette ottiche permettono di sondare il comportamento di +complessi proteici sottoposti a stimoli meccanici variabili, osservando +ad esempio come questi posssano modulare l'interazione tra due proteine +diverse. La teoria alla base del loro funzionamento è introdotta nella sezione \ref{sec:ot}. + +Quando sono combinate con tecniche ultraveloci per il posizionamento +delle trappole e il rilevamento degli spostamenti degli oggetti intrappolati le pinzette ottiche consentono la realizzazione di esperimenti di \emph{spettroscopia force-clamp}, approfonditi nella sezione \ref{sec:force_clamp}. + +Parallelamente la microscopia ottica ha permesso di descrivere i processi biologici con una precisione sempre maggiore, rendendo possibile la rilevazione e il tracciamento di singole molecole. In particolare +nell'ambito della microscopia di fluorescenza sono state sviluppate +tecniche per ricostruire immagini superando il \emph{limite di diffrazione}, per indurre la produzione di proteine fluorescenti grazie +all'ingegneria genetica, per rendere rilevabile il segnale di singoli fluorofori immobilizzati sopprimendo il rumore di quelli liberi in +soluzione. La teoria alla base di alcune di queste techniche è introdotta nella sezione \ref{sec:imaging}. + +Lo scopo di questa tesi è combinare un sistema di \emph{spettroscopia force-clamp} con un sistema di \emph{imaging di singola molecola} per l'esecuzione di misure in vitro simultanee e sincronizzate. + +In questo modo sarà possibile studiare, in un ambiente +controllato (proteine in soluzione e immobilizzate su microsfere), +il comportamento di proteine \emph{meccano-sensibili}, unendo alle +informazioni meccaniche quelle sulla dinamica di interazione +con altri fattori opportunamente marcarti presenti in soluzione. + + + +% Introduction on the importance of mechanotransduction + + + + %%%%%%%%% %%%%%%%%% %%%%%%%%% %%%%%%%%% %%%%%%%%% %%%%%%%%% %%%%%%%%% + + + +% between + +\section{Pinzette ottiche} +\label{sec:ot} + + +Le pinzette ottiche (o \textit{optical tweezers}, OT) sono strumenti che sfruttano la \emph{forza di radiazione} esercitata da un fascio laser gaussiano altamente focalizzato su materiali dielettrici, in modo da intrappolare e manipolare oggetti microscopici con una precisione sub-nanometrica. + +Questa tecnologia sfrutta il gradiente d'intensità di un fascio gaussiano focalizzato in prossimità del suo \textit{waist} e l'interazione tra il dipolo elettrico indotto nel materiale e il fascio. + +Arthur Ashkin fu, nel 1986, il primo a realizzare sperimentalmente delle pinzette ottiche, riuscendo a intrappolare microsfere sintetiche e batteri\cite{Ashkin:86}. Per questo risultato gli fu conferito il premio Nobel nel 2018, \emph{``per le pinzette ottiche e le loro applicazioni ai sistemi biologici''}. + +Per descrivere quantitativamente il funzionamento delle pinzette ottiche possiamo considerare l'interazione radiazione-materia nel limite di oggetti molto più piccoli della lunghezza d'onda della radiazione. + +In questo limite possiamo considerare il materiale interagente con la radiazione come un dipolo elettrico puntiforme, associato ad una polarizzabilità $\alpha$. Il vettore di polarizzazione nel dipolo puntiforme sarà quindi $\vec{p} = \alpha \vec{E}$. + +La forze esercitata su un dipolo elettrico puntiforme può essere ricavata a partire dalle \emph{legge di Lorentz}, ottenendo: + +$$ \vec{F} = + \left( \vec{p} \cdot \vec{\nabla} \right) \vec{E} + + \frac{d\vec{p}}{dt} \times \vec{B} +$$ + +Ovvero, una volta sostituito il vettore di polarizzazione: + +$$ \vec{F} = \alpha +\left[ + \left( \vec{E} \cdot \vec{\nabla} \right) \vec{E} + + \frac{d\vec{E}}{dt} \times \vec{B} +\right] +$$ + +E infine, tenendo conto delle \emph{equazioni di Maxwell} e dell'algebra dei vettori: + +\begin{equation} +\label{dipole_force} +\vec{F} += \alpha +\left[ + \frac{1}{2}\nabla E^2 + + \frac{d}{dt}\left(\vec{E} \times \vec{B}\right) +\right] +\end{equation} + +Questa ultima forma (equazione \ref{dipole_force}) ci permette di mettere in evidenza il termine $\frac{d}{dt}(\vec{E} \times \vec{B})$, ovvero la derivata temporale di una quantità oscillante alla stessa frequenza ottica del fascio laser (\SI{> 1e14}{\Hz}). + +Confrontando questo valore con la frequenza con cui riusciamo a campionare sperimentalmente il valore della forza risulta accurato considerare questa quantità costante, e quindi trascurare il secondo termine. + + +\section{Spettroscopia force-clamp} + diff --git a/chapters/2-methods.tex b/chapters/2-methods.tex new file mode 100644 index 0000000..b16855e --- /dev/null +++ b/chapters/2-methods.tex @@ -0,0 +1,152 @@ + %%%%%%%%% %%%%%%%%% %%%%%%%%% %%%%%%%%% %%%%%%%%% %%%%%%%%% %%%%%%%%% +\chapter{Metodi} + +In figure \label{fig:setup} è mostrato uno schema integrale dell'apparato realizzato. +Le componenti principali sono descritte nella sezione \ref{sec:setup}. +Successivamente, nella sezione \ref{sec:stabilization} è descritto il sistema di stabilizzazione meccanica introdotto per compensare lo spostamento +del campione dovuto a \textit{drift} termico e oscillazioni acustiche. +Nella sezione \ref{sec:calibration} è descritta nel dettaglio la procedura +usata per la calibrazione dei parametri delle trappole ottiche. + +L'esperimento tipico utilizzato per + +\section{Apparato} +\label{sec:setup} + +\begin{sidewaysfigure} + \includegraphics[width=1.0\linewidth]{images/Setup.pdf} + \caption{Caption} + \label{fig:setup} +\end{sidewaysfigure} + +\begin{sidewaystable} + \centering + \begin{tabular}{>{\tt}l l l >{\it}l l} + \toprule + Code & Type & Usage & Parameters & Product ID\\ + \midrule + \multicolumn{5}{l}{\it Fluorescence Excitation Path}\\ + F-Clean635 + & Bandpass Filter + & \SI{635}{\nm} laser clean-up + & $\lambda_{centr}: \SI{640}{\nm}, + bw: \SI{14}{\nm}$ + & FF01-640/14-25\\ + L-TS1a + & Plano-Convex Spherical Lens + & Telescope for \SI{635}{\nm} laser + & f: \SI{0}{\mm} + & \\ + L-TS1b + & & + & f: \SI{0}{\mm} + & \\ + L-TS2a + & + & Telescope for \SI{488}{\nm} laser + & f: \SI{0}{\mm} + & \\ + L-TS2b + & & + & f: \SI{0}{\mm} + & \\ + DIC-532 + & Single-edge Dichroic Filter + & Excitation beam multiplexing + & $\lambda_{edge}: \SI{532}{\nm}$ + & Di02-R532-25-D\\ + DIC-488 + & & + & $\lambda_{edge}: \SI{488}{\nm}$ + & Di02-R488-25-D\\ + L-TS3a + & Achromatic Doublet + & Telescope for comb. ex. lasers + & f: \SI{0}{\mm} + & \\ + L-TS3b + & & + & f: \SI{0}{\mm} + & \\ + L-Exc + & + & Excitation beam focusing + & f: \SI{0}{\mm} + & \\ + DIC-Fluor + & Quad-edge Dichroic Filter + & Excitation/Emission separation + & $\lambda_{edge}: \SIlist{405;488;532;635}{\nm}$ + & Di03-R405/488/532/635-t1 \\ + \multicolumn{5}{l}{\it Tweezers Path}\\ + L-TS4a + & Plano-Convex Spherical Lens + & Telescope matching isolator size + & f: \SI{0}{\mm} + & \\ + L-TS4b + & & + & f: \SI{0}{\mm} + & \\ + L-TS5a + & Plano-Convex Spherical Lens + & Telescope after isolator + & f: \SI{0}{\mm} + & \\ + L-TS5b + & & + & f: \SI{0}{\mm} + & \\ + L-Twz1 + & Plano-Convex Spherical Lens + & Telescope after isolator + & f: \SI{0}{\mm} + & \\ + L-Twz2 + & & + & f: \SI{0}{\mm} + & \\ + DIC-Twz1 + & Single-edge Dichroic Filter + & Tweezer beam insertion + & $\lambda_{edge}: \SI{1064}{\nm}$ + & \\ + \multicolumn{5}{l}{\it Imaging Path}\\ + L-Im1 + & Tube Lens + & Tube Lens + & f: \SI{0}{\mm} + & \\ + L-Im2 + & Imaging Lens + & Imaging Lens + & f: \SI{0}{\mm} + & \\ + DIC-BF + & Single-edge Dichroic Filter + & Fluorescence/Brightfield demux + & $\lambda_{edge}: \SI{705}{\nm}$ + & FF705-Di01-25x36\\ + F-BlockBF + & Shortpass Filter + & Residual brightfield suppression + & $\lambda_{cut}: \SI{700}{\nm}$ + & FESH0700\\ + F-Emiss + & Quad-band bandpass Filter + & Fluorophore emission filtering + & $\lambda_{centr}: \SIlist{446;510;581;703}{\nm}$ + & FF01-446/510/581/703\\ + F-BlockTwz + & Bandstop filter + & Fluorophore emission filtering + & $\lambda_{centr}: \SIlist{446;510;581;703}{\nm}$ + & FF01-446/510/581/703\\ + \end{tabular} + \caption{Caption} + \label{tab:my_label} + \end{sidewaystable} + + + \section{Stabilizzazione meccanica} + \label{sec:stabilization} \ No newline at end of file diff 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+ + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + diff --git a/main.tex b/main.tex new file mode 100644 index 0000000..a212bb4 --- /dev/null +++ b/main.tex @@ -0,0 +1,56 @@ +\documentclass[12pt,twoside,openright]{report} + + +% ===== PAGE LAYOUT +\usepackage[a4paper,width=150mm,top=25mm,bottom=25mm]{geometry} +\usepackage[utf8]{inputenc} +\usepackage{fancyhdr} + +% ===== LANGUAGE +\usepackage[italian]{babel} + +% ===== FIGURES AND CAPTIONS +\usepackage{graphicx} +\graphicspath{ {images/} } + +% ===== TABLES +\usepackage{booktabs} +\usepackage{rotating} + +% ===== TEXT FONTS +\usepackage{anyfontsize} +\usepackage{xcolor} +\usepackage{siunitx} +\sisetup{retain-unity-mantissa = false, + list-final-separator = {,}, + list-units = single} + +% ===== BIBLIOGRAPHY +\usepackage{cite} + +\title{Master Thesis} +\author{Lorenzo Zolfanelli} +\date{April 2020} + +\begin{document} + +\pagestyle{empty} +\input{titlepage} + +\newgeometry{width=150mm,top=25mm,bottom=25mm} +\pagestyle{fancy} +\setlength{\headheight}{15pt} + +\chapter*{Abstract} +\selectfont +Abstract goes here + +\tableofcontents + +\input{chapters/1-introduction} +\input{chapters/2-methods} +%\input{setup-fig/fig1} + +\bibliography{references.bib}{} +\bibliographystyle{plain} +\end{document} diff --git a/references.bib b/references.bib new file mode 100644 index 0000000..3140aa9 --- /dev/null +++ b/references.bib @@ -0,0 +1,16 @@ +@article{Ashkin:86, +author = {A. Ashkin and J. M. Dziedzic and J. E. Bjorkholm and Steven Chu}, +journal = {Opt. Lett.}, +keywords = {Laser beams; Optical trapping; Optical tweezers; Particle scattering; Radiation pressure; Scattering}, +number = {5}, +pages = {288--290}, +publisher = {OSA}, +title = {Observation of a single-beam gradient force optical trap for dielectric particles}, +volume = {11}, +month = {May}, +year = {1986}, +url = {http://ol.osa.org/abstract.cfm?URI=ol-11-5-288}, +doi = {10.1364/OL.11.000288}, +abstract = {Optical trapping of dielectric particles by a single-beam gradient force trap was demonstrated for the first reported time. This confirms the concept of negative light pressure due to the gradient force. Trapping was observed over the entire range of particle size from 10 $\mu$m to ~25 nm in water. Use of the new trap extends the size range of macroscopic particles accessible to optical trapping and manipulation well into the Rayleigh size regime. Application of this trapping principle to atom trapping is considered.}, +} + diff --git a/setup-fig/fig1.tex b/setup-fig/fig1.tex new file mode 100644 index 0000000..01ec068 --- /dev/null +++ b/setup-fig/fig1.tex @@ -0,0 +1,8 @@ +\begin{pspicture}[showgrid](0,-0.3)(3,3.3) +\pnode(0,2.5){A} +\pnode(2,2.5){B} +\pnode(2,1.5){C} +\mirror[labelangle=-45](A)(B)(C){M} +\optbox[position=start,labeloffset=0,labelref=relative](C)(B){Laser 1064 nm} +\drawbeam(A){1}{2} +\end{pspicture} \ No newline at end of file diff --git a/texlive/texlive.profile b/texlive/texlive.profile new file mode 100644 index 0000000..c03c68f --- /dev/null +++ b/texlive/texlive.profile @@ -0,0 +1,10 @@ +selected_scheme scheme-basic +TEXDIR /tmp/texlive +TEXMFCONFIG ~/.texlive/texmf-config +TEXMFHOME ~/texmf +TEXMFLOCAL /tmp/texlive/texmf-local +TEXMFSYSCONFIG /tmp/texlive/texmf-config +TEXMFSYSVAR /tmp/texlive/texmf/var +TEXMFVAR ~/.texlive/texmf-var +option_doc 0 +option_src 0 \ No newline at end of file diff --git a/texlive/texlive_install.sh b/texlive/texlive_install.sh new file mode 100644 index 0000000..094c6aa --- /dev/null +++ b/texlive/texlive_install.sh @@ -0,0 +1,33 @@ +#!/usr/bin/env sh + +# Originally from https://github.com/PHPirates/travis-ci-latex-pdf + +# This script is used for building LaTeX files using Travis +# A minimal current TL is installed adding only the packages that are +# required + +TEXLIVE_TAR_URL="http://mirror.ctan.org/systems/texlive/tlnet/install-tl-unx.tar.gz" + +# See if there is a cached version of TL available +export PATH=/tmp/texlive/bin/x86_64-linux:$PATH +if ! command -v texlua > /dev/null; then + # Obtain TeX Live + wget $TEXLIVE_TAR_URL + tar -xzf $(basename $TEXLIVE_TAR_URL) + # Install a minimal system + cd install-tl-20* + ./install-tl --profile=../texlive/texlive.profile + cd .. +fi + +# Just including texlua so the cache check above works +tlmgr install luatex + +# We specify the directory in which it is located texlive_packages +tlmgr install $(sed 's/\s*#.*//;/^\s*$/d' texlive/texlive_packages) + +# Keep no backups (not required, simply makes cache bigger) +tlmgr option -- autobackup 0 + +# Update the TL install but add nothing new +tlmgr update --self --all --no-auto-install \ No newline at end of file diff --git a/texlive/texlive_packages b/texlive/texlive_packages new file mode 100644 index 0000000..4400c33 --- /dev/null +++ b/texlive/texlive_packages @@ -0,0 +1,21 @@ +# In the case you have to install LaTeX packages manually, you can check +# https://www.ctan.org/pkg/some-package to see in which TeX Live package it is contained, +# or use tlmgr info some-package + +# Base tools to compile +texliveonfly latexmk + +# Collections +collection-langeuropean +collection-fontsrecommended + +# LuaTeX +luaotfload + +# PSTricks +#pst-arrow +#pst-3d +#pst-math + +# texliveonfly does not detect the following packages automatically +minted fvextra upquote lineno xstring framed caption \ No newline at end of file diff --git a/titlepage.tex b/titlepage.tex new file mode 100644 index 0000000..d1801aa --- /dev/null +++ b/titlepage.tex @@ -0,0 +1,74 @@ +\newenvironment{helv}{\fontfamily{qhv}\selectfont}{\par} + +\begin{titlepage} +%\newgeometry{left=20mm,right=15mm,top=15mm,bottom=15mm} +\begin{helv} + \begin{center} + \begin{minipage}{0.52\textwidth} + \includegraphics[width=\textwidth]{unifi} + \end{minipage} + \hfill + \begin{minipage}{0.4\textwidth} + \begin{flushright} + + \fontsize{14}{16}\selectfont + \textbf{\textcolor{gray}{ + Scuola di\\ + Scienze Matematiche\\ + Fisiche e Naturali\\ + }} + \vspace{12pt} + \fontsize{12}{14}\selectfont + Corso di Laurea Magistrale in\\ + Scienze Fisiche e Astrofisiche\\ + + \end{flushright} + \end{minipage} + \vfill + \end{center} + + \hspace*{22mm} + \begin{minipage}{0.8\textwidth} + \begin{flushleft} + \fontsize{36}{42}\selectfont + % ===== ENGLISH TITLE + \textbf{Titolo} + \vspace{36pt} + + % ===== ITALIAN TITLE + \textbf{Title.} + + + \vspace{3cm} + { + \fontsize{14}{0}\selectfont + \textbf{Relatore} + + \vspace{5pt} + \fontsize{16}{0}\selectfont + Marco Capitanio + + \vspace{15pt} + \fontsize{14}{0}\selectfont + \textbf{Correlatore} + + \vspace{5pt} + \fontsize{16}{0}\selectfont + Nome Cognome + + \vspace{15pt} + \fontsize{14}{0}\selectfont + \textbf{Candidato} + + \vspace{5pt} + \fontsize{16}{0}\selectfont + Lorenzo Zolfanelli + + \vspace{3cm} + \fontsize{14}{0}\selectfont + \textbf{Anno Accademico} ~ 2019/2020 + } + \end{flushleft} + \end{minipage} +\end{helv} +\end{titlepage} \ No newline at end of file