Life Science & Microscopy
Correct sample-, medium- and instrument-induced aberrations without folding the beam path.
Sharp at depth, in the microscope you already have
Every micrometre of depth costs signal.
The coverslip, the immersion medium and the tissue itself each distort the wavefront, and by a few hundred micrometres into a sample the structure that was crisp at the surface has washed into the background.
The Deformable Phase Plate puts it back. It is a transmissive corrector — refractive, in line, at the pupil plane — so it goes into an existing beam path without mirrors, relay optics or a redesign of the instrument around it. Two-photon, three-photon, light-sheet, confocal, widefield, single-molecule: the same part, in the same place, on commercial stands and custom builds alike.
Ways of integrating the DPP
five installations at partner labs
Commercial brightfield microscopePhaseform GmbH
Fluorescence, single-molecule setup with camera attachment PHITU Wien, Biophysics Research Unit · Dr. Mario BrameshuberTwo case studies
Two partner instruments, both corrected in line: a commercial two- and three-photon microscope at the DZNE in Bonn, and a custom light-sheet microscope at ICFO Barcelona running a Delta 7 in each of its two paths.
Deep in fixed brain, and inside a spheroid
Thorlabs Bergamo, two- and three-photon · DZNE Bonn, Light Microscopy FacilityThree-photon excitation is what you reach for when the tissue is too thick and too scattering for two-photon: a longer wavelength penetrates further, and the excitation volume is confined tightly enough that out-of-focus background all but disappears. The price is signal. Three-photon fluorescence scales with the cube of the intensity at the focus, so any aberration that spreads that focus comes back cubed — and several hundred micrometres into brain tissue there is no guide star to measure the wavefront against.
At the DZNE in Bonn the correction went into a commercial Thorlabs Bergamo, in a dual configuration: a Delta 7-10 in the illumination path and a Delta 7-20 in the detection path. Both are refractive and sit in line, so the Bergamo’s beam path was not folded or relayed to accommodate them. Phinden does the rest sensorlessly — it optimizes on the image the microscope is already producing, driving the instrument through its own ThorImage software, so nothing has to be measured that a scattering sample cannot give.
The frame below is fixed mouse brain at 420 µm, three-photon at 1300 nm — the depth at which scattering and aberration together normally end the experiment. Both halves are the same field, acquired the same way. Only one of them has the correction on.
The same instrument, the same correction loop, one floor down in excitation order: an organic spheroid at two-photon, where the aberration to remove is the sample’s own refractive index rather than several hundred micrometres of brain.
A Delta 7 in each path of a light-sheet microscope
custom light-sheet setup · SLN Lab, ICFO BarcelonaAt the SLN Lab at ICFO Barcelona, pig sperm cells are imaged in a custom light-sheet microscope: low-magnification Nikon objectives on both sides, the sample inside a tube with water around it. What the experiment is after is the fine morphology of the tail — and the tail is the first thing to go, thin enough that the aberrations of the system and the sample, together with the limited depth of field of the setup, obscure it.
So both halves of the optical system were corrected at once. A Delta 7-10 went into the illumination path behind the 10× objective, in a 30 mm cage with the objective screwed straight onto it; a Delta 7-20 went under the 16× detection objective, mounted horizontally on top of the Z-stage. Neither path was folded or relayed to make room, and the full mechanical integration into the existing setup took under an hour. Phinden found the correction sensorlessly, from the images the microscope was already producing. The illumination-side unit was not only correcting, either: it was used to shape the light sheet itself, holding contrast across a wider field.
Let’s start with your sample
What is your specimen doing to the wavefront?
This is the one that matters. A sample is not a uniform slab: nuclei, lipid droplets, vasculature, myelin and cell walls each carry their own refractive index, and light crossing them arrives at the objective with a wavefront that no lens was designed for. It changes with the specimen, with the field, and as you move through it. That is the aberration adaptive optics exists for — and the one no better objective will fix.
Is the immersion medium the one your objective was computed for?
Silicone oil in place of oil, a mounting medium that changed with the protocol, a chamber nobody specified: each mismatch appears as spherical aberration that grows with depth. Refocusing does not touch it. The correction runs out to the 6th-order spherical term.
How deep into the sample are you working?
Clearing is never perfect. The clearing medium never quite matches the tissue interior, so a depth-dependent spherical term rides on top of whatever vasculature, dense nuclei and myelin remnants add locally. The wavefront leaving a millimetre-deep sample is not the one that entered it.
Do you want the aberration — on purpose?
Not every experiment wants a perfect focus. Three-dimensional single-molecule localization reads depth out of a deliberately engineered point-spread function — and the 63 electrodes that take astigmatism out can just as well put it in, calibrated and repeatable, with no phase mask to swap.
Each of these has been corrected on a working instrument at a partner lab. Further results on the PHI page and in the microscopy application note.
Your instrument has a wavefront signature of its own too — tube lens, filter cube, dichroic, a stressed mount. Measured once on beads under the correct immersion, it becomes the baseline the sample correction starts from instead of an unknown mixed in with everything else.
If any of them describes your setup: how much of your sample are you still not seeing?
Your application is not on this list?
Let us build a customized DPP product based on your requirements to realize your vision of a novel way to apply adaptive optics.