Schedule

Introduction to CMSS2023

Organising Committee

Correlative imaging –using ions and single molecules

Professor Dr. Aleksandra Radenovic

From the plethora of correlative imaging modalities, SR techniques were most frequently combined with electron microscopy to provide protein-ultrastructure relationships at nanometer-scale resolution. At the other forefront of methods in development, scanning probe microscopy techniques aim to capture nanoscale topographical dynamic changes of cells under physiological conditions. Professor Radenovic will present a combined SICM-SR, which avoids membrane deformation and unlocks long-term monitoring of the biological processes.

Flexibility of correlative light and electron Microscopy (CLEM): from virus to organ

Dr. Jemima Burden

Correlative multimodal imaging can help find the needle in the haystack and answer more questions that either technique alone. Dr Jemima Burden will give an overview of the breadth and flexibility of the approach and how it can help researchers address questions across scales, from the level of a single virus, to understanding cell behaviour within a whole organ.

Multiscale imaging of microbial cellular complexes

Prof. Dr. Martin Pilhofer

Prof. Dr. Pilhofer will discuss integrated approaches to identify and characterize cellular complexes by (cryo-) light microscopy, cryo-electron-tomography and single particle cryo-EM. The methods will be illustrated by examples including bacterial contractile injection systems, as well as eukaryotic and archaeal filamentous assemblies. 

Title to be confirmed 

Dr. Arne Seitz

X-ray view on cellular and organismal morphogenesis

Dr. Venera Weinhardt

X-ray imaging methods have a unique advantage over other methods, as it provides high spatial resolution and high penetration power for hydrated specimens with minimal sample preparation prior to imaging. Dr. Venera Weinard will present an overview of ongoing developments in deploying X-ray imaging methods to capture morphogenesis at diverse scales of bioimaging.

Multi-scale and multi-modal correlative imaging of neuronal circuits

Dr. Adrian Wanner

During learning, experience-driven modifications of synaptic connections are thought to store information in the connectivity pattern—the wiring diagram—to optimize responses to future inputs. Detailed analyses of neuronal connectivity at synaptic resolution are therefore critical to understand neuronal computations and memory. Dr. Adrian Wanner will present the advantages of using correlative imaging modalities to interpret the connectomics patterns with unprecedented resolution. 

Advances in integrated fluorescence microscopy for cryo-CLEM

Sponsor talk: Thermo Fisher Scientific

Experimental considerations for structure localisation using 3D cryo-CLEM

Dr. Herman Fung

3D cryo-CLEM can help us localise structures of interest in situ during cryo-ET sample preparation, data acquisition and interpretation. In this lecture, we will examine the important considerations for each step of the workflow, from sample design to imaging and registration. As well, we will touch on complementary solutions such as on-lamella CLEM, with the end goal of building a pragmatic workflow that maximises throughput and accuracy towards the biological subject at hand.

Sample preparation of bulk specimens for cryo-FIB and        cryo-ET

Dr. Bettina Zens

In this lecture, Dr. Zens will discuss the steps necessary to prepare complex specimens such as tissues or 3D cell culture samples for cryo-ET: A combination of High-Pressure Freezing (HPF), cryo-CLEM, and cryo-FIB milling using the lift-out technique allows the acquisition of in situ data on bulk samples. Several examples will be used to show the challenges in each step and strategies to overcome them.

CLEM methods for tissue in 2D and 3D

Dr. José Maria Mateos Melero

During this lecture, Dr. Mateos Melero will present methods they apply at their Facility to correlate protein expression with subcellular details of the tissue and workflows to identify the same cell/organelle in the 3D space between light and electron microscopes.

Group 1: CLEM workshop

DCI and UNIL

Workshop, covering the steps of CLEM pipeline. We will demonstrate plunge-freezing, cryo-LM, FIBSEM and cryo-TEM for cryo-electron tomography. 

Group 2: 

BIOP

Correlative soft X-ray and fluorescence tomography for single-cell quantitative analysis 

Dr. Valentina Loconte

SXT takes advantage of the naturally occurring, differential X-ray absorption of the carbon-rich compounds in each organelle.  When correlated with light microscopy, it becomes a powerful tool to correlate local changes in mesoscale molecular densities to the functional output of complex chemical processes of living cells. Dr. Valentina  will present a detailed outlook of these techniques to answer long-standing questions in biology. 

X-Ray nanotomography of bulk materials using ptychography 

Dr. Ana Diaz 

X-ray ptychography (XRP) and its 3D counterpart, ptychographic X-ray computed tomography (PXCT) have a huge potential in bioimaging. XRP and PXCT are relatively modern scanning coherent diffraction imaging techniques delivering high spatial resolutions (e.g., routinely <50 nm) and are limited neither by focusing optics (beam size) nor by the scanning step size, and applicable to extended sample volumes up to ≈100 μm diameter. In this talk, Dr. Ana Diaz will present detailed steps involved in ptyhographic imaging and the future outlook of its application to tackle complex biological questions. 

Hyperspectral imaging of soft matter with soft X-ray STXM

Dr. Benjamin Watts

Hyperspectral Soft X-ray radiation offers decisive advantages over conventional imaging techniques as it is sensitive to chemical bonds and oxidation states. In his talk, Dr. Benjamiin Watts  will present the protocols of setting up soft X-ray STXM samples and future outlook on using these methods for bioimaging. 

Correlative solutions in 3D: combining X-ray microscopy and FIB tomography

Sponsor Talk: ZEISS

Group 2: CLEM workshop

DCI and UNIL

Workshop, covering the steps of CLEM pipeline. We will demonstrate plunge-freezing, cryo-LM, FIBSEM and cryo-TEM for cryo-electron tomography. 

Group 1: 

BIOP

Scanning Near-field Optical Microscopy: a good (and one of the first) example of correlative microscopy

Dr. Sergey Sekatskii, EPFL 

In his talk, Dr. Sekatski will introduce us to one of the first examples of correlative microscopy in the scanning probe world. Scanning Near-Field Optical Microscopy is a very fundamental technique combining scanning probes with lasers to get the topography and optical properties of the samples under study. He will dwell on the fundamentals of these techniques and discuss their current applications in biology.

SICM: Scanning Ion-Conductance Microscopy

Samuel Mendes Leitão 

In this talk, Samuel will discuss about time-resolved scanning ion conductance microscopy (SICM), a method for non-invasive live cell imaging at the nanoscale. He will present the development of a SICM microscope from concept to prototype to study processes on cell membranes in 3D, such as morphological changes in cancer cells and bacteria-host cell infection. He will also show the advantage of combining SICM with optical fluorescence imaging for additional insights into molecular activity involved in membrane processes. To finalize the talk, he will present a new nanopore method for single-molecule detection using SICM, characterizing DNA molecules with correlative fluorescence measurements. 

Nanoendoscopy AFM: a window into the cell 

Dr. Marcos Penedo Garcia, EPFL 

In this lecture, Dr. Penedo Garcia will give a very interesting insight into how scanning probes can be used to look into the cell and not just it's surface. He will present his work of combining fluorescence microscopy with Atomic Force microscopy to image the inside of the cell with sharp probes fabricated using Focused Ion Beam-Scanning Electron microscopy, a good example of applications of diverse microscopic modalities.

Correlative AFM approaches Workshop

Instrumentation challenges in AFM-SEM combination, Advances in cantilever fabrication for correlative approaches and Applications of correlative AFM-LM imaging

Ten Core Principles of Image Analysis

Dr. Daniel Sage

Correlative cryo-fluorescence electron tomography to examine dopaminergic neurons

Dr. Lycas Mathhew Domenic

In his talk, Dr Lycas will discuss Cryo-EM tomography as a method and how it can be used to reveal the ultrastructure of synapses and has the potential for in situ protein structure identification. However, it is limited by its need to have relevant biological samples on specific substrates in order to allow for imaging to work. Here, Dr. Lycas will demonstrate the growth of dopaminergic neuron co-cultures over the cryo-EM grids, and following genetic labeling strategies, the identification of dopaminergic neurons with cryo-fluorescence. With this identification, they are able to target dopaminergic synapses with cryo-EM tomography and to capture detailed tomograms of these locations. Using this data they are able to identify in situ protein structure as well as pharmacological changes to organelle distribution. 

Examination