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Stampwell

powered by the Mount(n) technology


Designed by Gaëlle Recher

"The Mount(n) technology is the results of my expertise in imaging a wide range of 3D thick samples. I navigated through many communities (gathered around a biological model), and each time learnt new tricks to mount samples for microscopy experiments. The Mount(n) technology consists in imprinting an array of wells into an hydrogel (typically agarose), each well receiving a 3D sample (embryos of any species, organoids, tissue biopsies...). The design of the wells prevents the object from moving when the plate or dish is moved, does not apply any mechanical constrain to the sample, and paradoxically also enables the sample to keep growing inside. In a single dish, tens of samples can be imaged with any microscope, for as long as necessary."  

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    EXAMPLES OF RESULTS

     APPLICATIONS

      TECHNOLOGY FEATURES

      METHODS

     PUBLICATIONS

  EXAMPLES OF RESULTS

The first stamp to imprint arrays of wells in hydrogels and to mount 3D biological samples for parallelized microscopy experiments. Stampwell ensures the sample stillness, long-term medium-throughput and automatized multi-position image acquisition for a significant timesaving.

 
 
Imprinted well in agarose
Soaked in fluorescein, negative rendering
Image credit: (c)Gaëlle Recher - Bordeaux
 
 
Cell-filled alginate capsule in the well, imaged in z
Notice that the focus moving through the well height is easily visible (lines in focus Vs out of focus)
Image credit: (c)Gaëlle Recher - Bordeaux
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Imprinted wells
Filled with a single cell-loaded alginate capsules; corresponding labels imaged with a stereomicroscope and stitched. Gel pad is made of Agarose 2%. Bar: 2mm.
Image credit: (c)Gaëlle Recher - Bordeaux
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Zebrafish larva
In rectangle shape wells - bar=1mm (projection of Z-stack - confocal microscope)
Image credit: (c)Gaëlle Recher - Bordeaux
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Cancer cells co-culture
In alginate capsule - bar = 100um
Image credit: (c)Gaëlle Recher, Simon Latour - Bordeaux
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hiPSCs cyst in alginate capsule
bar = 500um
Image credit: (c)Gaëlle Recher, Maxime Feyeux - Bordeaux

  APPLICATIONS

Domains:

  • Developmental biology
  • Cell biology
  • Tissue engineering
  • Morphogenesis
  • High/medium throughput - many samples on the same dish
  • Imaging (all microscopes) & culturing 3D samples
  • Suitable for both live or fixed samples

Techniques:

  • Organoids

  • Spheroids

  • Embryos

  • Tissues explants

  TECHNOLOGY FEATURES

Medium/high throughput: 70 objects can be loaded with one stamp

Automatized multi-position image acquisition: Stampwell provides a constraint-free but stable and parallelized immobilization of the samples allowing automatized multi-position image acquisition

Long-term imaging: Because the sample is not embedded in the agarose pad but just laying on the bottom of a V-shaped well, it can freely grow within the well, enabeling culture and image aquisition over days and weeks

  METHODS

Storage conditions

  • Room temperature 

Protocol of use  

  • Quickly pour liquid agarose (ideally: 2% normal gelling temperature agarose, or other hydrogel of your choice, it has to be stiff enough once reticulated)

  • Quickly place the stamp: put one side of the stamp in contact with the bottom of the dish, and then tilt it cautiously until the whole surface of the stamp is in contact with the dish, to avoid the formation of air bubbles (protips: if doing in a 6-well plate, think about aligning the arrays in each well to better automatise image aquisition with the microscope software)

  • Reticulate the hydrogel (for agarose, cool down, you can place the whole thing at 4°C to speed up the process)

  • To remove the stamp: a neat result is obtained by passing a scalpel blade all around the stamp to make sure it is well detached from the gel, remove the stamp by tilting a bit at first and pulling firmly, if possible with a non-wounding object, hold the gel pad to prevent it to detach from the holder

  • If you've prepared your gel from water, it is recommended to preincubate the whole gel with culture medium a few hours at the desired temperature to homogenize the solution in the hydrogel, and rince once

  • Load the samples in each well (you can use either an embryologist mouth pipet if you know how to or a yellow tip) (protip: before pipeting your favourite sample, prerinse your tip or pipet with serum to avoid it to stick to it)

  • Go to the microscope (and if possible, automatise the acquisition, the spacing between wells is standard)

  PUBLICATIONS

Alessandri K, Andrique L, Feyeux M, Bikfalvi A, Nassoy P, Recher G. All-in-one 3D printed microscopy chamber for multidimensional imaging, the UniverSlide. Sci Rep. 2017 Feb 10;7:42378. https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5301227/