Crowd-funding a high-NA multi-immersion objective lens

We have designed a promising high-NA multi-immersion all-immersion objective lens that has applications for both cleared tissue imaging and live-cell imaging.  We would love to make this objective generally available, but we need the help of the research community.

The initial design was completed just prior to the COVID pandemic and there are interested researchers, but we are stuck in a chicken-and-egg situation 1.5 years later. For custom optics like this there are large fixed costs for both design and manufacturing, so to get a reasonable per-unit price you need to make a lot of them.  But making a lot of them is a large investment that could be a loss if they don’t sell.

We were recently in a similar situation with the “King Snout” objective lens, but enough scientists placed pre-orders that ASI was comfortable proceeding with a production run. We’d like to replicate the same model with this all-immersion objective lens. Consider it a crowdfunding appeal; ASI is willing to bear significant financial risk, but only if there are enough customers who agree to purchase the resulting product.

So here’s the offer: ASI will proceed with a production run if we receive POs for seven of these objectives by end-July 2021, with expected delivery in November 2021.  We will offer a special price of $25k for any orders received by July 31. Update: We did not meet our goal of getting commitments for 7 of these, however the interested leading scientists are still interested so we are keeping a lookout for anyone else who would be willing to pre-order these.Leading scientists have already promised to purchase 3 of them, so we need another 4 customers to get us over the hump.  Any production run will be for at least 15 objectives, so there will be some left for future orders.  But the scientists who pre-order will not only be the first to get their hands on them, but will also further the entire field by helping to make this objective lens a reality.

We are happy to share CAD and Zemax black box models.  Please contact me at jon@asiimaging.com for more information or to request a formal quote.

Here are the main specifications:

  • Magnification / NA in BABB : “56x” / NA 1.24
  • Magnification / NA in CLARITY: “52x” / NA 1.15
  • Magnification / NA in water: “48x” / NA 1.05
  • Working distance: 3.4 mm
  • Optical / mechanical angle: 52° / 57°
  • Field of view: 0.5 mm
  • AR coatings: 400 – 1300 nm
  • Plan-corrected, apochromatic, optimized for visible
  • 83 mm parfocal distance, M25x0.75 threads

What will this objective be good for? I’m glad you asked! There are at least 3 categories of applications:

    1. Cleared tissue imaging
    2. Single-objective light sheet
    3. High-NA light sheet (e.g. lattice light sheet)

Obviously this objective lens will be an excellent choice for any application involving high-resolution cleared tissue imaging.  It will have the highest NA — and hence best resolution — of any objective for imaging cleared tissue.  It can be used in any of the various media for cleared tissue, spanning the refractive index (RI) range from water to BABB or ethyl cinnamate.  Like our other multi-immersion objectives it features a curved first surface which allows it to be used in any dipping medium without a correction collar.  We expect it to be resistant to harsh organic chemicals like DBE and BABB like our other multi-immersion objectives.

Single-objective light sheet and remote refocus rely on creating a remote 3D image of the sample through a carefully-constructed optical relay, which requires adjusting the system magnification to match the sample.  This magnification change happens automatically if an all-immersion objective is used as the primary objective.  Hence this objective presents the opportunity to construct a versatile single-objective microscope system that can be used for various media, as well as having exceptional working distance and 52° optical angle.  Even though it is a dipping objective, it is possible to use a coverslip at the sample and correct for it at the remote image (by using a coverslip-corrected objective without a coverslip for O2).

Finally, this objective lens will be useful for any sort of high-NA live imaging including lattice light sheet.  The original Nikon 25x/1.1 W lens used for lattice light sheet has many disadvantages, but it has excellent “mechanical efficiency” or large NA for the amount of room it takes up near the sample.  Our new objective fits into the same space near the sample as the Nikon 25x/1.1 W, but it has significantly more working distance (3.4 vs. 2.0 mm), is 8 mm longer (83 vs. 75 mm parfocal distance), has magnification more suited to high-resolution imaging, and a flat field.  The penalty is a slightly reduced numerical aperture, NA 1.05 in water instead of NA 1.1 (5% worse lateral resolution and 11% worse axial resolution).  An under-appreciated fact is that the exact working distance (WD) of most dipping/immersion objectives changes as the exact RI of the medium changes — including as the temperature changes — owing to refraction at the objective/medium interface.  Thus long-term light sheet imaging usually requires continual adjustment of either the objective or light sheet position.  However, all three of ASI’s multi-immersion objectives has the exact same WD regardless of medium RI, simplifying experiments.

Here are some links for those interested in digging deeper:

Help us make King Snout a reality

We have received a number of inquiries about the AMS-AGY v2 “King Snout” objective.  We would love to make it generally available, but we need the help of the research community.

This objective has been stuck in a chicken-and-egg situation.  ASI needs some orders to feel confident proceeding with a production run.  (If we don’t sell very many we could end up with expensive glass sitting on the shelf and lose money, whereas the unit price is prohibitive for a tiny production run.)   However, if ASI is waffling then researchers aren’t inclined to send orders.

We came up with the following plan: ASI will proceed with a production run if we receive POs for at least four of the AMS-AGY v2 objectives by mid-January 2021, with expected delivery in April 2021.  We will offer a special price of $25k for any orders received by mid-January.  Afterwards, the expected price is $29.5k.  Please contact me at jon@asiimaging.com if you would like a formal quotation. (Clarification: a production run will be for 10+ objectives, so if a production run happens there will be some left for future orders). [Update 20-Jan-21: We have met the order threshold and a production run is in process.]

Both AMS-AGY objectives — v1 “Snouty” and v2 “King Snout” — make it easy to implement high-NA single-objective light-sheet (SOLS) microscopes using a variety of primary objective lenses.  High-NA SOLS combines the simple sample-mounting of traditional (single-objective) microscopes with the rapid and gentle imaging that light sheet provides, without compromising resolution as earlier approaches did.  The AMS-AGY objectives are used as the tertiary objective in high-NA SOLS.  They were invented by Calico employees Alfred Millett-Sikking and Andrew G. York, are manufactured by Special Optics, and are distributed by ASI.  The v1 objective has been commercially available for almost a year now with a price of $15k, and this push is to make the v2 become commercially available as well.


Relative sizes of v2 and v1 objectives. From Box 1 of preprint at https://doi.org/10.1101/2020.09.22.309229.

A larger field of view (FOV) is v2’s raison d’être.  The v1 has FOV of ∅150 μm (diffraction-limited) and ∅250 μm (usable).  The area of v2’s FOV is 9-fold larger at ∅450 μm (diffraction-limited) and ∅750 μm (usable).  Besides applications where simultaneously imaging different parts of a larger sample is required, v2 enables faster acquisitions of static samples by filling more of the camera at a time, e.g. reducing the number of tiles required.

The benefit of v2 depends on the application.  For imaging cells with high-magnification primary objective lenses (e.g. 100x/1.35 silicone), the v2 likely confers no benefit and v1 is more cost-effective.  For imaging larger samples with modest magnifications (e.g. 20x-40x primary objective), the v2 offers tremendously improved throughput.  Importantly, v2 is a strict upgrade; if you can afford the extra expense there is no technical reason to prefer the v1.

The potential of the AMS-AGY v2 objective is beautifully demonstrated in a recent preprint from Chan Zuckerberg Biohub by Bin Yang, Loïc Royer, and collaborators.  They used a single-objective light sheet microscope to monitor the development of a zebrafish tail.  Their home-built microscope had a 20x/1.0 water primary objective and the imaged volume was 1000 μm x 550 μm x 250 μm (they used stage scanning to achieve 1000 μm but the Y and Z dimensions are instantaneous).  They implemented several clever ideas, and central to their technical achievements was their use of a (prototype) AMS-AGY v2 objective to achieve an effective NA ≈ 1.0.  This is significantly higher NA (read better resolution) than ever previously demonstrated over such a large field of view with a single-objective light sheet microscope.

Some links for those interested in digging deeper are below.  Please let me know if you have questions and/or can help us make this objective a commercial reality by ordering one in the next few months.


From left to right: original AMS-AGY v1 objective (OD 23 mm), prototype AMS-AGY v2 objective, and a familiar object for scale. Photo courtesy Alfred Millett-Sikking.

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Brand-New MS-2000 Non Flat Top Stage with Controller for Sale

We are offering brand-new MS-2000 Non Flat Top Stages for $7.500.00 for the following Microscopes:

  • TE/TI2000
  • IX71/81. It won’t work with IX73/83
  • IX70
  • DMI6000. It won’t work with DMi8
  • AV2000
  • TE300

Description:
Specifically designed to provide a high resolution, and highly repeatable, means of controlling the X and Y position of the microscope stage. All axes derive their precise control through the use of closed-loop DC servomotors employing high-resolution rotary encoders for positioning feedback. The MS-2000 XY stage utilizes crossed-roller slides, a high-precision lead screw, and zero-backlash miniature geared DC servomotors for smooth and accurate motion. The Z-axis drive also uses ASI’s proven line of closed-loop motor drives, each custom fitted to the microscope. The microprocessor-controlled MS-2000 control unit provides for RS-232 and USB communication with a host computer.

Specifications:

  • XY axis range of travel: 120 mm x 110 mm
  • XY axis resolution (encoder step): 22 mm
  • XY axis RMS repeatability: <700 nm
  • XY axis maximum velocity: 7 mm/sec
  • Max Recommended Load: 5kg

For more information go to our SPECIAL OFFERS page.