Corporate Banner
Satellite Banner
Scientific Community
Become a Member | Sign in
Home>News>This Article

Scientists at the London Research Institute of CRUK Use the Linkam CMS196

Published: Thursday, November 07, 2013
Last Updated: Thursday, November 07, 2013
Bookmark and Share
Scientists use CMS196 stage for the imaging of mammalian cells with Cryo-CLXM microscopy.

Linkam Scientific Instruments has reported on the use of their innovative CMS196 cryo stage for the study of mammalian cells at the London Research Institute, Cancer Research UK.

For mammalian cells to remain in a healthy state, they require constant renewal of their components. The process of disposing of old components is known as 'autophagy', which stems from the Greek words auto "self" and phagein "eat".

This process involves the formation of a double-membrane structure called an autophagosome, which engulfs old or dysfunctional organelles and then fuses with lysosomes, where they are broken down to recycle the constituent molecules.

Autophagy is increased when cells are starved, and plays a fundamental role in a large number of cellular processes, including development, immunity, neurodegeneration and cancer.

In a recent publication in the journal, Ultramicroscopy (Duke et al., 2013), Dr Lucy Collinson (LRI Electron Microscopy Unit), in collaboration with Dr Sharon Tooze (LRI Secretory Pathways Lab), imaged forming autophagosomes in whole mammalian cells.

The structures are particularly difficult to capture in cells prepared for electron microscopy, so they are now using a powerful new technique called cryo-soft X-ray tomography, cryo-SXT, working with Dr Liz Duke at the Diamond Light Source synchrotron.

This allows whole mammalian cells to be imaged as close to the living state as possible. The cells are grown on tiny gold grids and plunged into liquid ethane to preserve the cells in the frozen state.

In order to find the autophagosomes within the cells, they are labelled with green fluorescent protein (GFP). The fluorescent autophagosomes are then located using a technique called correlative cryo-fluorescence and cryo-soft x-ray microscopy (cryo-CLXM).

Cryo-fluorescence microscopy is performed using the Linkam CMS196 stage prior to the cells being transported in cryo-containers to synchrotrons in Oxfordshire, Berlin and Barcelona for imaging. One of the major advantages of this new correlative approach is that the CMS196 stage allows the cells to be screened for quality and protein localization in the research laboratory before actually travelling to the synchrotron, which is critical in terms of cost and efficiency.

The combination of cryo-fluorescence microscopy and cryo-SXT allows scientists to link the functionality of proteins to their near native-state structure. This should find wide applications in cell biology studies of health and disease.

The Linkam CMS196 stage was designed specifically to solve the problem of how to get vitrified EM grids from the fluorescence microscope into the cryo-TEM without devitrification and contamination through condensation. The stage has been optimized optically to enable the use of high NA lenses.

Up to 3 grids can be loaded into a specially designed cassette for transportation from the plunge freezer to the upright fluorescent microscope. The cassette is then easily loaded onto the viewing bridge using special manipulation tools.

The sample viewing chamber is perfectly dry and below -180ºC while the sample bridge itself is at -196°C. The grids can be quickly and efficiently scanned using a 100X 0.75NA lens and manipulated using high precision micrometers. The cassette is then simply manipulated back into the transportation device and is then transported to the cryo-TEM under liquid nitrogen.

Further Information
Access to this exclusive content is for Technology Networks Premium members only.

Join Technology Networks Premium for free access to:

  • Exclusive articles
  • Presentations from international conferences
  • Over 2,600+ scientific posters on ePosters
  • More than 3,800+ scientific videos on LabTube
  • 35 community eNewsletters

Sign In

Forgotten your details? Click Here
If you are not a member you can join here

*Please note: By logging into you agree to accept the use of cookies. To find out more about the cookies we use and how to delete them, see our privacy policy.

Scientific News
Lung Repair and Regeneration Gene Discovered
New role for hedgehog gene offers better understanding of lung disease.
3 Ways Viruses Have Changed Science for the Better
Viruses are really good at what they do, and we’ve been able to harness their skills to learn about – and potentially improve – human health in several ways.
Mixed Up Cell Transportation Key Piece of ALS and Dementia Puzzle
Researchers from the University of Toronto are one step closer to solving this incredibly complex puzzle, offering hope for treatment.
New Gene Therapy for Vision Loss From a Mitochondrial Disease
NIH-funded study shows success in targeting mitochondrial DNA in mice.
Five New Genetic Variants Linked to Brain Cancer Identified
The biggest ever study of DNA from people with glioma – the most common form of brain cancer – has discovered five new genetic variants associated with the disease.
Predictive Model for Breast Cancer Progression
Biomedical engineers have demonstrated a proof-of-principle technique that could give women and their oncologists more personalized information to help them choose options for treating breast cancer.
Fatty Liver Disease and Scarring Have Strong Genetic Component
Researchers say that hepatic fibrosis, which involves scarring of the liver that can result in dysfunction and, in severe cases, cirrhosis and cancer, may be as much a consequence of genetics as environmental factors.
Specific Variations in RNA Splicing Linked to Breast Cancer
Researchers have identified cellular changes that may play a role in converting normal breast cells into tumors. Targeting these changes could potentially lead to therapies for some forms of breast cancer.
Finding Links and Missing Genes
A catalogue of large-scale genetic changes around the world.
Scientists Test New Gene Therapy for Vision Loss from a Mitochondrial Disease
NIH-funded study shows success in targeting mitochondrial DNA in mice.
Skyscraper Banner

Skyscraper Banner
Go to LabTube
Go to eposters
Access to the latest scientific news
Exclusive articles
Upload and share your posters on ePosters
Latest presentations and webinars
View a library of 1,800+ scientific and medical posters
2,600+ scientific and medical posters
A library of 2,500+ scientific videos on LabTube
3,800+ scientific videos