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Jon Daniels

@jsdaniel02.bsky.social

440 followers 530 following 42 posts

Scientific instrument developer, focused on #lightsheet and other #microscopy 🔬 at ASI

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Jon Daniels @jsdaniel02.bsky.social · 9d

BTW these objectives generally work for water/ExM though not specifically optimized for it. For instance the 54-12-8 -- NA 0.64 in water -- was used as the illumination objective for ASLM imaging of pan-ExM samples in www.biorxiv.org/content/10.1...

Pan-ASLM: a high-resolution and large field-of-view light sheet microscope for Expansion Microscopy

Expansion microscopy, a super-resolution fluorescence microscopy technique in which samples are expanded up to ∼8,000 times (after 20-fold expansion) their original volume, places high demands on the ...

www.biorxiv.org

Jon Daniels @jsdaniel02.bsky.social · 9d

of course

Jon Daniels @jsdaniel02.bsky.social · 9d

In the past this (and the glycerol one) was available for purchase only with Olympus/Evident confocal systems, not to homebuilders. Not sure if they have changed their policy.

Jon Daniels @jsdaniel02.bsky.social · 9d

for cleared tissue see these four objectives with different NA/WD/FOV tradeoffs: asiimaging.com/docs/multi-i...

multi-immersion_objectives [Applied Scientific Instrumentation]

Jon Daniels @jsdaniel02.bsky.social · 9d

Note this would be a dipping objective and basically a regular microscope objective lens, compared to the specialized ExaSPIM lenses which are awesome but unconventional optically.

Jon Daniels @jsdaniel02.bsky.social · 9d

I have some thoughts in this direction... I'm thinking NA 0.75 or NA 0.8 with >10mm WD.

I'm hoping to talk with researchers at Seeing Is Believing and SfN (or by email/DM) and then start the detailed design.

Please reach out to me [email protected].

Jon Daniels @jsdaniel02.bsky.social · 19d

Here is what it will look like:

Jon Daniels @jsdaniel02.bsky.social · 19d

Jon Daniels @jsdaniel02.bsky.social · 20d

OK not sure why it's not posting as either png or jpg...

Jon Daniels @jsdaniel02.bsky.social · 20d

Jon Daniels @jsdaniel02.bsky.social · 20d

I like Empress but I don't want to be the arbiter of nicknames.

Official PN is 57-14-7 and it will be colloquially "AMS-AGY v3" on our website.

It's 10mm shorter than v2. OD is flared near the threads: 30mm OD for most and 37mm OD near the threads, whereas v2 was 35mm OD all the way.

Jon Daniels @jsdaniel02.bsky.social · Jul 9

Details here: bsky.app/profile/jsda...

Jon Daniels @jsdaniel02.bsky.social · Jul 9

Production has started for a new #Snoutscope objective, AMS-AGY v3. It has significantly larger FOV and some other tweaks from v1 and v2. Currently accepting pre-orders. The price will need to increase modestly after deliveries start late October.

comparison table for v2 and v3

Jon Daniels @jsdaniel02.bsky.social · Jul 9

Tagging @amsikking.bsky.social @andrewgyork.bsky.social @tanner-fadero.bsky.social @retof.bsky.social @loicaroyer.bsky.social and I'm sure I'm missing more but hopefully this will make the rounds...

Jon Daniels @jsdaniel02.bsky.social · Jul 9

High-res cellular imaging uses O1 > 1.0 with O2 being 40x/0.95 02. Short O2 WD means the grind has to sacrifice FOV in the depth axis, but FOV in the other direction (parallel the coverslip) is still 600um.

For larger samples e.g. zebrafish the grind position is wider to capture the entire FOV.

Jon Daniels @jsdaniel02.bsky.social · Jul 9

4. 55° grind so compatible with any-immersion concept.

5. Two different options for grind position: one for situations where O2 is NA ~0.95 and another for NA ~0.8. This one requires a bit more explanation.

Jon Daniels @jsdaniel02.bsky.social · Jul 9

A few nerdy details:

1. Generally better imaging quality especially for those pushing the FOV. I can send plots to those interested.

2. The image plane is a few microns off the glass tip to mitigate problem with dust.

3. Improved capture of high-angle rays despite slightly reduced NA of 0.99.

Jon Daniels @jsdaniel02.bsky.social · Jul 9

Production has started for a new #Snoutscope objective, AMS-AGY v3. It has significantly larger FOV and some other tweaks from v1 and v2. Currently accepting pre-orders. The price will need to increase modestly after deliveries start late October.

comparison table for v2 and v3

Jon Daniels @jsdaniel02.bsky.social · Jul 1

OK, makes sense. There is a loss in perfect 3D focusing away from the native focal plane described in the Botcherby paper and seemingly there in practice. Can that also be derived from this analytic approach?

Jon Daniels @jsdaniel02.bsky.social · Jul 1

Really cool! Could this be extended to focal planes other than the "native" one?

Jon Daniels @jsdaniel02.bsky.social · May 19

The equation from the paper is equivalent to 4.676 pixels per "Rayleigh diffraction limit square" (i.e. change that one number and my calculation lines up)

Even if you don't use all the "pixels" in such an objective, there is speed/brightness benefit to having such a large NA over such a large FOV.

Jon Daniels @jsdaniel02.bsky.social · May 19

Can you explain a bit more how you calculate it? I'm probably missing a factor.

FOV is π/4*(7200^2) in units of μm^2

Rayleigh diffraction limit is 0.61*λ/NA in units of μm

Can support 4 pixels per (diffraction limit)^2 (is this right?)

At 500nm: π/4*(7200^2)/(0.61*0.5/0.5)^2*4 = 438 MPix

Jon Daniels @jsdaniel02.bsky.social · May 15

I'm excited to be able to be in Zurich for the mesoSPIM symposium. This is a really fantastic project.

Jon Daniels @jsdaniel02.bsky.social · Apr 18

We're about to start sending quotes, should be delivered late summer / early fall. Happy to discuss via email.

Jon Daniels @jsdaniel02.bsky.social · Apr 18

v3 is coming which will functionally replace v2, and should be modestly better in a few respects. So no reason to make more v2, at least for now.

Jon Daniels @jsdaniel02.bsky.social · Mar 17

I'll have to think about that one