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Automated Inverted Microscope for Life Science Research Leica DMI4000 B

Archived Product
Replaced by Leica DMi8

Circuit board in LED incident light: glare prevents a clear view of the solder points

Glare-free viewing is achieved by using the polarization set. There is a noticeable reduction in eyestrain.

Visual Inspection with the Stereomicroscope

Polarization Set for Fatigue-Free Work

Every photographer knows the value of a polarization filter for absorbing disturbing light reflections – whether from water surfaces or shiny metals. But light reflections can be a serious problem in industry, too. For example, reflecting incident light makes it considerably more difficult to inspect circuit boards, electronic components and other metal workpieces. It is almost impossible to inspect a sample swiftly and accurately if light reflections prevent a clear view.

A simple and efficient solution is to use a polarization set on the stereomicroscope. The filter can be steplessly regulated and individually adjusted for each sample. By rotating the filter through 90 degrees, its effect can be varied between 0 and 100 per cent. The surface of the sample immediately appears clear, vivid and full of contrast. Any material defects or other visible flaws can be identified quickly and confidently. For employees that have to work at the microscope for long periods of time, this means significantly less eye strain and improved working conditions. For  the company, it means substantial improvements in efficiency and enhanced work quality.

Many Leica stereomicroscopes can be retrofitted to offer these benefits: The polarization set is an optional accessory for the Leica ring lights LED3000 RL and LED5000 RL. An investment that pays off very quickly.

(A) Ultrathin section of P. tricornutum expressing Pex10-GFP in Epon without antibody labeling. The boxed area is shown in (B) at higher magnification and illustrates two peroxisomes in proximity to the nucleus, the golgi and the plastid. CW, cell wall; G

(A) Ultrathin section of P. tricornutum expressing Pex10-GFP in Epon without antibody labeling. The boxed area is shown in (B) at higher magnification and illustrates two peroxisomes in proximity to the nucleus, the golgi and the plastid. CW, cell wall; G, golgi apparatus; Mt, mitochondrium; Ne, nuclear envelope; Nu, nucleus; P, peroxisome; Pl, plastid; V, vacuole. Scale bars represent 2 μm (A), 500 nm (B).

Targeting of peroxisomal matrix proteins in the diatom Phaeodactylum tricornutum

P. tricornutum cells expressing different types of GFP fusion proteins were harvested via centrifugation at 1,500xg and cryoimmobilized by high-pressure freezing on gold carriers

Sample Preparation:
P. tricornutum cells expressing different types of GFP fusion proteins were harvested via centrifugation at 1,500xg and cryoimmobilized by high-pressure freezing on gold carriers (Leica EM PACT2, Leica Microsystems GmbH, Vienna, Austria). Subsequently, the cells were freeze-substituted with acetone in combination with 2% OsO4, 0.25% uranyl acetate and 5% H2O. Freeze substitution was carried out in the automated Leica EM AFS2 unit (Leica Microsystems GmbH, Vienna, Austria) at -90°C for 4 h, -60°C for 8 h, -30°C for 8 h and then held at 0°C for at least 3 h ...

For more details please see:

http://www.plosone.org/article/info%3Adoi%2F10.1371%2Fjournal.pone.0025316

Courtesy of:
Dr. Kathrin Bolte and Dr. Andreas Klingl
Zellbiologie & Elektronenmikroskopie
AG Prof. Uwe-G. Maier
Philipps Universität Marburg, Marburg
The website of the EM facility is:
http://www.synmikro.com/de/startseite/core-facilities/elektronenmikroskopie.html

<media 50974 - external-link-new-window "Opens external link in new window">Download the Application Note</media>

Ultrastructure of Arabidopsis thaliana primary root cells in PIN1pro::PIN1=GFP;bex5-1 treated for 1h with 50 uM BFA.

Sample Preparation of Arabidopsis thaliana root tips cells

by Riet De Rycke, resp. DMBR-PSB Transmission Electron Microscopy-Core Facility. Department for Molecular Biomedical Research, Plant Systems Biology, Gent

This Application Note describes the preparation method "Electron Microscopy of high pressure frozen and freeze substituted Arabidopsis thaliana root tips cells" using Leica EM AFS2.

Arabidopsis thaliana roots (mutant PIN1pro:PIN1-GFP;bex5-1) were excised, immersed in 20% (w/v) BSA and frozen immediately in a high-pressure freezer (EM PACT; Leica Microsystems, Vienna, Austria).

Freeze substitution was carried out using a Leica EM AFS2 (Leica Microsystems) in dry acetone containing 1% ddH2O, 1% OsO4 and 0.5% glutaraldehyde over a 4-days period as follows:

-90°C for 24 hours, 2°C per hour increase for 15 hours, -60°C for 24 hours, 2°C per hour increase for 15 hours, and -30°C for 24 hours. Samples were then washed 3 times in pure acetone between -30°C and 0°C and slowly warmed up to 4°C, infiltrated stepwise over 3 days at 4°C in Spurr’s resin and embedded in capsules. The polymerization was performed at 70 °C for 16 h.

Ultrathin sections were made using an ultramicrotome (Leica EM UC6) and post-stained in in a Leica EM AC20 for 40 min in uranyl acetate at 20 °C and for 10 min in lead citrate at 20 °C. Grids were viewed with a JEM 1010 transmission electron microscope (JEOL, Tokyo, Japan) operating at 80 kV using Image Plate Technology from Ditabis (Pforzheim, Germany).

<media 50501 _blank external-link-new-window "Application Note: Electron Microscopy of high pressure frozen and freeze substituted Arabidopsis thaliana root tips cells">Download Application Note "Electron Microscopy of high pressure frozen and freeze substituted Arabidopsis thaliana root tips cells" (pdf, 3,73MB)</media>

High Magnification Sperm inspection (IMSI)

IMSI is an advanced ICSI method descibed by Bartoov et al. (in Journal of Andrology 2002, 23).

Prior to sperm injection the morphology of the sperm cell is evaluated with high magnification DIC microscopy.
Leica Microsystems offers an aplication optimized system called the Leica "Big S". This has been developped in close cooperation with the lab of Prof. S. Viville, Strassbourg, France. Leica AM6000 with "Big S" has been installed in multiple IVF labs.

Discover the fine structure of living sperm with Interference contrast

  • Superb objective HCX PL FLUOTAR 100x/ 1.30 oil
  • Continuous increase and decrease of magnification and your sperm! Without changing objectives
  • Magnify sperm –approx. 16000 x on a 19" monitor
  • Ergonomic and – Simple handling
  • Leica DFC 290 digital camera – Brilliant life image with high refreshing rate
  • Leica evaluation software – Measure and qualify sperm morphology
  • Adaptable to all Leica DMI
  • Leica AM6000 - Big S association – Workflow optimised system to control microscope and micromanipulator simultaneously
  • ICSI and IMSI on one station

λ2 plot of defined region of interest

White Light Laser – the Ultimate Source for Confocal Microscopy

The perfect light source for confocal microscopes in biomedical applications has sufficient intensity, tunable color and is pulsed for use in lifetime fluorescence.

Furthermore, it should be a pulsed source for lifetime fluorescence experiments. Such a source has been invented: the white light laser

Rolf T. Borlinghaus, Dr. 1 , Lioba Kuschel, Dr. 1
1
Leica Microsystems, Wetzlar, Germany

Read article: White Light Laser – the Ultimate Source for Confocal Microscopy

 

FCS – Principle of Fluorescence Correlation Spectroscopy

Fluorescence Correlation Spectroscopy: The Femtoliter Test Tube – System Calibration and In Vitro Applications

Fluorescence correlation spectroscopy ( FCS ) measures fluctuations of fluorescence intensity in a sub-femtolitre volume to detect such parameters as the diffusion time, number of molecules or dark states of fluorescently labeled molecules.

The technique was independently developed by Watt Webb and Rudolf Rigler during the early 1970s. The term FCS was coined by the Webb lab.

Read article: Fluorescence Correlation Spectroscopy: The Femtoliter Test Tube – System Calibration and
In Vitro
Applications

Recruitment of HP1b to UV-damaged chromatin is influenced by the histone acetylation state

How to Study Protein Recruitment to DNA Lesions by a Combination of UV Laser and White Light Laser

Understanding how DNA lesions are optimally repaired is of functional significance, especially from the view of genome karyotype stability.

DNA lesions are recognized by several proteins in a hierarchical manner: First, an appearance of double strand breaks (DSBs) initiate recruitment of primary protein complexes, such as MRE11-RAD50-NBS1, which are responsible for the activation of a DNA damage-related serine/threonine protein kinase called ataxia telangiectasia mutated (ATM)...

Read article: How to Study Protein Recruitment to DNA Lesions by a Combination of UV Laser and White Light LaserHow to Study Protein Recruitment to DNA Lesions by a Combination of UV Laser and White Light Laser

CARS image of cellulose fibers. The fibers are visualized through the C–H vibrations of the polyglucan chains in cellulose.

Imaging Characteristic Vibrational Contrast of Molecules

Coherent anti-Stokes Raman scattering (CARS) microscopy is a technique that generates images based on the vibrational signatures of molecules.

This imaging methods does not require labeling, yet molecular specific information can be obtained from a range of important bio-molecular compounds.

Read article: Imaging Characteristic Vibrational Contrast of Molecules

Rayon anisotropy: CARS anisotropy image of rayon fibers. The anisotropy was determined at the 2880 cm–1 methylene stretching mode of rayon. The brightness in the image corresponds to the degree of orientation of the methylene mode in the fiber.

Good Vibrations - Perspectives of CARS Microscopy in Life Science Research

The ability to see the cell’s molecular machinery at work has contributed immensely to our understanding of cellular functioning.

In recent years, new molecular imaging techniques, such as coherent anti-Stokes Raman scattering microscopy (CARS), have been developed for rapid vibrational imaging of living cells. The intrinsic molecular vibrations leave specific fingerprints in the vibrational spectrum.

Eric Olaf Potma, Ph.D. 1 , Bernd Sägmüller, Dr. 2
1
University of California, Irvine (UCI), Department of Chemistry, USA, 2Leica Microsystems, Wetzlar, Germany

Read article: Good Vibrations - Perspectives of CARS Microscopy in Life Science

In vivo imaging of food moth (Plodia interpunctella). Surface of a larval silk gland, 10x magnification.

CARS Microscopy – an Introduction

CARS overcomes the drawbacks of conventional staining methods by the intrinsic characteristics of the method.

CARS does not require labeling because it is highly specific to molecular compounds which are based on vibrational contrast and chemical selectivity. The crucial advantage of this method is that the sample remains almost unaffected

Stefanie Degenhartt, Dr.
Leica Microsystems, Wetzlar, Germany

Read article: CARS Microscopy – an Introduction