Lyncee Tec DHM T-series

Description

The Digital Holographic Microscopy (DHM®) is a patented technology. It records, with a digital camera, holograms produced by the interference between the beam transmitted through the sample, and a reference beam generated inside of the microscope. The holograms are processed numerically to reconstruct a 3D optical map of the specimen.

The DHM® T-series is a transmission configured Digital Holographic Microscope and optical profilometer for applications in life science and material science: a unique experimental set-up enabling characterization of light transmitting samples. It enables non-toxic, label-free quantitative measurements of individual living cells and cultures up to confluence.

The vertical calibration of DHM is intrinsically defined by the laser wavelength. It provides high accuracy and reproducible data and measures with interferometric resolution, i.e. a subnanometric vertical resolution, and a lateral resolution limited by the choice of microscope objective. Thanks to advanced numerical processing of the recorded hologram, sharp focus can be performed simultaneously or after measurement, as a post-processing without manually adjusting the height of the sample.

Two configurations of Transmission DHM are available:

T1000 models: Configured with a single wavelength and are the ideal tool for studying simple living cells as well as measuring transparent material samples with smooth surfaces.
T2100 models: Configured for measuring simultaneously at two wavelengths, extending measurement capability.

Applications

Biological Imaging & Life Science: Label-free biological microscopy, time-lapse, multi-well plate screening, diagnostics, and investigation of unexplored biological processes.
Biological Process Interpretation: Quantitative phase measurements can be interpreted in terms of channel activity, cell viability, intracellular concentration, and morphology changes.
Multimodal Imaging: Combined with an optional fluorescence module for simultaneous DHM and fluorescence measurements, decreasing the number of necessary fluorescence labelling and enhancing understanding of cellular mechanisms. It can also be combined with an electrophysiological setup.
Material Science & Innovative Materials/Devices: Measurement of sample optical topography, thickness variability, size and location of internal structures and defects, refractive indices and concentration, and birefringence.
Fast Dynamical Measurement Applications:
- Micro-optics
- Micro-fluidics
- Stress and constraints analysis
- Liquid Crystal Displays (LCD)
- Biophysics
- Wettability of coatings and structures
- Fluids and gas dynamics
- Dissolution and crystallization
- Particle velocimetry

Time sequence of 3D optical map of quantitative phase measurement , limited by camera rate : division of HeLa cells

Specifications

System
DHM models	T1000	T2100
Number of laser sources	1	2
Operating wavelength (± 1.0 nm)	666 nm	666 nm, 794 nm
Laser wavelength stability	0.01 nm/°C at 675 nm
Sample stage	Manual or motorized XYZ stages 114 mm x 76 mm x 38 mm travel range
Objectives	Magnification 1.25x to 100x, standard, high NA, long working distance, water/oil immersion
Objective & condenser turret	6 positions
Computer	Workstation with latest multicore Intel® processor, high performance graphic card, optimized and configured for DHM with screen min 21inch and mouse
Software	Proprietary Koala software based on C++ and .NET Additional optional software modules available for advanced analysis
Data compatibility	Measurement data recorded in bin format, exportable in .txt format, recorded and reconstructed images exportable in .tif format or .txt array
Performance
Measurement mode	Single wavelength at 666 nm	Short synthetic wavelength at 8 μm4
DHM models	T1000, T2100	T2100
Accuracy 1[nm]	1.04	1.0 / 5.0 4*
Vertical resolution 2[nm]	2.04	2.0 / 10.0 4*
Repeatability 3[nm]	0.024	0.02 / 0.05 4*
Vertical measuring range	up to 500 μm4	up to 500 μm4
Max. height of steps with sharp edges6	up to 1.0 μm4 up to 3.5 μm5	up to 7.0 μm4 up to 22 μm5
Vertical calibration	Determined by interferometric optical filter, ±0.1 nm
Acquisition time	Standard: 500 μs (optional 10 μs)
Acquisition rate	Standard: 190 fps (1024 x 1024 pixels). (optional up to 100'000 fps).
Reconstruction rate	Up to 25 fps 1024 x 1024 pixels hologram (data analysis dependent ). (optional up to 60 fps )
Lateral resolution	Objective dependent, down to 300 nm **
Field of view	Objective dependent, from 66 μm x 66 μm up to 5 mm x 5 mm **
Working distance	Objective dependent, from 0.3 to 18 mm **
Digital focusing range	Up to 50x depth of field (objective dependent)
Sample illumination	Down to 1 μW/cm2

Power requirements
Input voltage	85-260 VAC – 50/60 Hz
Power requirements	max. 250 W
Dimensions & weight
Dimensions (L x W x H)	600 x 350 x 500 mm
Weight	30 kg

1 As demonstrated by taking the temporal standard deviation on 1 pixel over 30 measurements
2 Defined as twice the accuracy
3 As demonstrated by taking the one sigma Rq value of 30 repeatability measurements without sample
4 Converted value for measurements in air and with sample refractive index n = 1.5
5 Converted value for measurements in water and with sample refractive index n = 1.5
6 Depends on the laser source(s) and operating wavelength(s)

* With / Without single wavelength mapping
** Objectives specifications on www.lynceetec.com/microscope-objectives

Options

Objectives with extra-LWD, cover-glass correction, for immersion, etc.
Motorized stage for automation and stitching
Remote TCP/IP module for automation and remote control of DHM
Stroboscopic unit for MEMS analysis
Dipping tip for immersion measurement in well plate and open chambers
Fluorescence module to combine DHM with epifluorescence measurements
High-speed camera to extend measurement capabilities