Lastek Principals

Digital Holographic Microscopy (DHM®) quantitative phase images record the morphology (shape) and content (hemoglobin content) of Red Blood Cells and provide an important clinical readout on the health status of RBC without label as demonstrated in this latest publication (see below) in the International Journal of Molecular Sciences.

Image above: Non-perturbing observation of flowing RBC with DHM® (20x magnification)

Unique advantages of DHM®
  • Morphology and intracellular content (hemoglobin) measurements
  • 194 recorded images per second: fast dynamics
  • Millisecond to multi-days continuous recording: time-lapse
  • Label-free, non-invasive: no cell perturbation
  • Automated microscope: high-content/throughput screening

 

Lyncée Tec DHM® T1000 for live cell Quantitative Phase Imaging

A complete solution from live cell environment to reporting

Lyncée delivers turn-key systems that include:

  • a modular and evolutive inverted DHM® T microscope
  • sample environmental control
  • multi-modal imaging add-ons
  • motorized samples stage for automation and screening
  • software with automated workflows, data analysis and reporting

The modular and evolutive DHM®-T fits any budget

The DHM® T-100 is the simplest of Lyncée inverted microscope offering an ideal cost effective tool for routine and non-perturbing cell culture monitoring at single and population level. It is equipped with a single fixed objective, that can be selected from large choice of magnification, numerical aperture, and coverslide corrections.

The DHM® T-100  can then later by upgraded to a DHM® T-1000 that support multiple objectives operation.

 

Recent paper:

Image- and Fluorescence-Based Test Shows Oxidant-Dependent Damages in Red Blood Cells and Enables Screening of Potential Protective Molecules

by Manon Bardyn 1,Jérôme Allard 1,2,David Crettaz 1,Benjamin Rappaz 3,Gerardo Turcatti 3,Jean-Daniel Tissot 4 andMichel Prudent 1,4,5,*'
 
Author to whom correspondence should be addressed.
Academic Editor: Anastasios G. Kriebardis
Int. J. Mol. Sci. 2021, 22(8), 4293; https://doi.org/10.3390/ijms22084293
Received: 28 February 2021 / Revised: 9 April 2021 / Accepted: 15 April 2021 / Published: 20 April 2021
(This article belongs to the Special Issue In Sickness and in Health: Erythrocyte Responses to Stress and Aging)

Abstract

An increase of oxygen saturation within blood bags and metabolic dysregulation occur during storage of red blood cells (RBCs). It leads to the gradual exhaustion of RBC antioxidant protective system and, consequently, to a deleterious state of oxidative stress that plays a major role in the apparition of the so-called storage lesions. The present study describes the use of a test (called TSOX) based on fluorescence and label-free morphology readouts to simply and quickly evaluate the oxidant and antioxidant properties of various compounds in controlled conditions. Here, TSOX was applied to RBCs treated with four antioxidants (ascorbic acid, uric acid, trolox and resveratrol) and three oxidants (AAPH, diamide and H2O2) at different concentrations. Two complementary readouts were chosen: first, where ROS generation was quantified using DCFH-DA fluorescent probe, and second, based on digital holographic microscopy that measures morphology alterations. All oxidants produced an increase of fluorescence, whereas H2O2 did not visibly impact the RBC morphology. Significant protection was observed in three out of four of the added molecules. Of note, resveratrol induced diamond-shape “Tirocytes”. The assay design was selected to be flexible, as well as compatible with high-throughput screening. In future experiments, the TSOX will serve to screen chemical libraries and probe molecules that could be added to the additive solution for RBCs storage. 

Description

An increase of oxygen saturation within blood bags and metabolic dysregulation occur during storage of red blood cells (RBCs). It leads to the gradual exhaustion of RBC antioxidant protective system and, consequently, to a deleterious state of oxidative stress that plays a major role in the apparition of the so-called storage lesions.

Material and methods

The screening assay was conducted on an automated DHM-T1000 with a motorized stage for High-content/high-throughput screening applications.

Digital Holographic Microscopy is used to quantify the morphological alterations induced by three oxidants (AAPH, diamide and H2O2) and the protective effect of four antioxidants molecules (ascorbic acid, uric acid, trolox and resveratrol)  at different concentrations.

Results

This approach allowed to identify a specific shape of RBC called “Tirocytes” and open the way to large screens of molecules on RBC to improve their storage health.

 

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Metrology

Principals: Metrology