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

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

Join For Free

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 3,500+ scientific posters on ePosters
  • More than 5,000+ 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 TechnologyNetworks.com 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
New Therapeutic Target for Crohn’s Disease
A promising new target for drugs that treat IBD has been identified along with a possible biomarker for IBD severity.
Uncovering Water Bear Resilience
A protein identified in water bears can protect DNA of human cells from lethal doses of radiation damage.
Using Stem Cells to Grow a 3D Lung-in-a-Dish
Researchers have created 3D lung-like tissue from lung-derived stem cells. The tissue can be used to study lung diseases.
Reprogramming Lymph Nodes to Fight MS
Bioengineers work to reprogram lymph node function to fight multiple sclerosis.
Puttng Cells Through Their Paces
An obstacle course for human lung cells could be the answer for better testing the effectiveness of potential new drugs.
Inherited Heart Condition Breakthrough
Using stem cells, scientists have created a specific heart condition model, yeilding insights into unexpected disease mechanisms.
Genetic Tug of War Before Cells Decide Fate
Researchers report that as developing blood cells are triggered by genetic signals firing on and off, a 'tug of war' occurs.
Origin of Cultured Cells: Not Where You Think
Study shows cultured cells from decades-old cell line does not originate from the patient it was claimed to derive from.
Hope for Zika Treatment Found in Drug Screening
Johns Hopkins researchers join collaborative group to screen 6,000 existing drugs in hopes of finding treatments for Zika Virus infection.
Worms Point Way Toward Viral Strategies
Rice University wins NIH grant to study how nematodes handle gastrointestinal viruses.
Skyscraper Banner

SELECTBIO Market Reports
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
3,500+ scientific and medical posters
A library of 2,500+ scientific videos on LabTube
5,000+ scientific videos
Close
Premium CrownJOIN TECHNOLOGY NETWORKS PREMIUM FOR FREE!