Transmission configured Digital Holographic Microscope (DHM®) for applications in life science andmaterial science: a unique experimental set-up enabling characterization of light transmitting samples.

DHM® - T series

Please visit www.lynceetec.com and contact us for more information and examples of applications.

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

Transmission DHM®

DIGITAL HOLOGRAPHIC MICROSCOPY

Label-free biological microscopy

DHM® enables non-toxic quantitative measurements of individual living cells and cultures up to confluence :

• Time-lapse • Multi-wellplatescreening • Diagnostic

Transmission configured optical profilometer

The transmission DHM® enables the measurement of multiple sample characteristics :

• Sampleopticaltopography • Thicknessvariability • Sizeandlocationofinternalstructuresanddefects • Refractiveindicesandconcentration • Birefringence

Investigate innovative materials and devices

Thebenefitswithfastdynamicalmeasurementhave been demonstrated in numerous applications :

• Micro-optics • Micro-fluidics • Stressandconstraintsanalysis • LiquidCrystalDisplays(LCD) • Biophysics • Wettabilityofcoatingsandstructures • Fluidsandgasdynamics • Dissolutionandcrystallization • Particlevelocimetry

Investigate unexplored biological processes

The quantitative phase measurement of DHM® can be interpreted in terms of many underlying biological processes :

• Channelactivity • Cellviability • Intracellularconcentration • Morphologychanges

Perform multimodal imaging

An optional module enables simultaneous DHM® and fluorescence measurements :

• CorrelateDHM® measurements withwellknownmeasurementprotocols • UsetheDHMtodecreasethenumber of necessary fluorescence labelling • Enhanceunderstandingofcellularmechanism

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DHM®

TheDigitalHolographicMicroscopy(DHM®)isapatentedtechnology.Itrecords,withadigitalcamera,holograms producedbytheinterferencebetweenthebeamtransmittedthroughthesample,andareferencebeamgeneratedinside of the microscope. The holograms are processed numerically to reconstruct a 3D optical map of the specimen.

The vertical calibration of DHM® is intrinsically defined by the laser wavelength. It provides high accuracy andreproducibledataandmeasureswithinterferometricresolution,i.e.asubnanometricverticalresolution,andalateralresolution limited by the choice of microscope objective.

Thankstoadvancednumericalprocessingoftherecordedhologram,sharpfocuscanbeperformedsimultaneouslyoraftermeasurement,asapost-processingwithoutmanuallyadjustingtheheightofthesample.

time

DHM-Tcombinedwithanelectrophysiologicalsetup

Represented byInnovation Park, Batiment A1015 LausanneSwitzerlandinfo@lynceetec.comwww.lynceetec.comDHM-T - 0814 / Printed in Switzerland / © Lyncée Tec SA 2014 / All rights reserved

Two configurations of Transmission DHM® are available, differentiated by the number of wavelengths.

> T1000 models are 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 are configured for measuring simultaneously at two wavelengths, extending measurement capability

DHM® systemsarecompatiblewithalargechoiceofoptions

• Objectiveswithextra-LWD,cover-glasscorrection,forimmersion,etc. • Motorizedstageforautomationandstitching • RemoteTCP/IPmoduleforautomationandremotecontrolofDHM • StroboscopicunitforMEMSanalysis • Dippingtipforimmersionmeasurementinwellplateandopenchambers • FluorescencemoduletocombineDHMwithepifluorescencemeasurements • High-speedcameratoextendmeasurementcapabilities

1Asdemonstratedbytakingthetemporalstandarddeviationon1pixelover30measurements 2Definedastwicetheaccuracy 3AsdemonstratedbytakingtheonesigmaRqvalueof30repeatabilitymeasurementswithoutsample 4Convertedvalueformeasurementsinairandwithsamplerefractiveindexn=1.5 5Convertedvalueformeasurementsinwaterandwithsamplerefractiveindexn=1.5 6Dependsonthelasersource(s)andoperatingwavelength(s)

*With/Withoutsinglewavelengthmapping

**Objectivesspecificationsonwww.lynceetec.com/microscope-objectives

Technical specifications

System

DHM models T1000 T2100

Number of laser sources 1 2

Operatingwavelength(±1.0nm) 666 nm 666 nm, 794 nm

Laserwavelengthstability 0.01nm/°Cat666nm

Sample stage ManualormotorizedXYZstages114mmx76mmx38mmtravelrange

Objectives Magnification1.25xto100x,standard,highNA,longworkingdistance,water/oilimmersion

Objective & condenser turret 6 positions

Computer DellworkstationwithlatestmulticoreIntel®processor,highperformancegraphiccard, optimizedandconfiguredforDHMwithscreenmin21inchandmouse

Software ProprietaryKoalasoftwarebasedonC++and.NETAdditionaloptionalsoftwaremodulesavailableforadvancedanalysis

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 Singlewavelengthat666nm Shortsyntheticwavelengthat8μm4

DHM models T1000,T2100 T2100

Accuracy 1 [nm] 1.04 1.0/5.04 *

Vertical resolution 2 [nm] 2.04 2.0/10.04 *

Repeatability 3 [nm] 0.024 0.02/0.054 *

Vertical measuring range upto500μm4 upto500μm4

Max.heightofstepswithsharpedges6 upto1.0μm4

upto3.5μm5upto7.0μm4

upto22μm5

Vertical calibration Determinedbyinterferometricopticalfilter,±0.1nm

Acquisition time Standard:500μs(optional10μs)

Acquisition rate Standard:30fps(1024x1024pixels).(optionalupto1000fps).

Reconstruction rate Upto25fps1024x1024pixelshologram(dataanalysisdependent).(optionalupto60fps)

Lateral resolution Objectivedependent,downto300nm**

Fieldofview Objectivedependent,from66μmx66μmupto5mmx5mm**

Workingdistance Objectivedependent,from0.3to18mm**

Digital focusing range Upto50xdepthoffield(objectivedependent)

Sample illumination Downto1μW/cm2

Powerrequirements

Input voltage 85-260VAC–50/60Hz

Powerrequirements max.250W

Dimensions&weight

Dimensions(LxWxH) 600x350x500mm

Weight 30kg

Transmission DHM®combinedwithaFluorescencemodule