PhiOptics Quantitative Phase Contrast Microscope

PhiOptics Quantitative Phase Contrast Microscope

– The quantitative phase imaging (QPI)/spatial light interference microscope (SLIM)

 

The microscope is used to visualize and quantify specimens, e.g. cells: shapes, height/volume, refractive index, dry mass, growth and transport. The SLIM module includes a spatial light modulator, needed to control the phase delays, and a camera to acquire the images for each phase delay. The quantitative phase image can be seamlessly overlaid with the epifluorescence image.

 

The microscope

The Axio Observer.Z1 is an inverted microscope with a touch pad and a mechanical stage. With the add-on SLIM module from PhiOptics, SLIM /QPI) is realized.

 

Focus drives

  • Motorized objective nosepiece (z-step size 10 nm)

 

Objectives (for phase contrast)

 

Magnification NA Type Immersion Coverslip WD (mm)
10x 0.3 Ph1 EC Plan-Neofluar Air - / 0.17 5.2
20x 0.8 Ph2 Plan-Apochromat Air 0.17 0.55
40x 0.75 Ph2 Plan-Neofluar Air 0.17 0.71
63x 1.40 Ph3 Plan-Apochromat Water Oil 0.10

 

Condenser lens

NA 0.55, air, working distance 26 mm, with phase rings

 

Light for epi-fluorescence and transmitted light

  • X-Cite 120Q, 120W Hg vapor short arc lamp (bulb lifetime 2000h) with liquid light guide.
  • Halogen lamp: 12V – 100W

 

Filter cubes for epi-fluorescence

 

Name Excitation filter Emission filter Dichroic
Filter Set 49/DAPI ~365 nm BP 445/50 395 nm
Filter Set 10/AF 588 BP 450 - 490 nm BP 515 - 565 nm 510 nm
Filter Set 43/CY3 BP 550/25 nm BO 605/70 590 nm 570 nm

 

Camera

Hamamatsu Orca Flash 4 v2 sCMOS, 4.2 MPix, 100 fps at max resolution

 

Software on the computer

CellVista from PhiOptics with modules for acquiring

  • Z-stacks
  • Tiles
  • Time series