ORDER & DELIVERY
How do I order products?
The Gri3D® plates with standard microwell sizes can be directly purchased from the webshop where you can proceed to online payment. It is also possible to simply request a quote by sending an email to [email protected] with your needs. In this case, a quote will be sent to you by email and the order can be placed accordingly. Learn more about the standard options here.
In the case of custom Gri3D® plate, the order can only be made by requesting a quote by sending an email to [email protected] with your needs. A quote will be sent to you by email and the order can be placed accordingly. Learn more about the customization options here.
Sample plates are also available to order if you want to first test which microwell size is more convenient for your application (only available for standard microwell sizes). In this case, you can place your sample plates order here or request one by sending an email to [email protected]. There is an order limitation of maximum 3 sample plates. Learn more about the sample plates here.
I need a quote for my institution, how do I request one?
You can request a quote by sending an email to [email protected] with your needs.
What are the accepted payment methods?
We accept the following online payment methods: PayPal, credit card or bank transfer.
How and when will my order be delivered?
We ship worldwide using standard room temperature courier service. Upon receipt of your order, we ship within 2-3 weeks. Accounting for transit time delivery, it should take approximately three weeks for standard orders. For custom orders, lead time is 4-5 weeks.
Refer to the Terms & Conditions for more details.
Can my order be delivered to a different address than my billing address?
Yes, you can specify your shipping address and your billing address when making your order.
To which countries does SUN bioscience ship orders?
We ship our products worldwide.
How can I cancel my order?
If you wish to cancel an order prior to the delivery, contact [email protected] .
What are the shipping costs?
Shipping costs are 33 CHF for national shipments in Switzerland and 55 CHF for international shipments.
I haven’t received my order, what should I do?
If you are still waiting for your order after the estimated delivery time, please send an email to [email protected] with your name and order number and a member of our team will get back to you.
GRI3D® PRODUCT SPECIFICATIONS
What is the shelf-life of Gri3D® and how should I store them?
Upon receipt, please store the plates at 4°C where they are stable for at least 6 months.
Do I need any special equipment to use Gri3D®?
Gri3D® products are SLAS-standard and are compatible with standard laboratory equipment such as multichannel pipets or high-content imagers.
Is Gri3D® automatable?
Yes. We have established automated organoid workflows including hydrogel equilibration, cell seeding, medium exchange and compound exposure with liquid handlers.
What are the plate dimensions?
96 wellplate: 127.90 x 85.60 x 14.45 mm (l x w x h).
24 wellplate: 127.90 x 85.60 x 19.00 mm (l x w x h).
Technical drawings are available upon request at [email protected] .
What are the recommended well volumes?
96 wellplate: 50 μl seeding chamber + 150 μl pipetting port.
24 wellplate: 200 μl seeding chamber + 800 μl pipetting port.
What material is Gri3D® made of?
Top plate: polystyrene.
Hydrogel microwell: poly-ethylene glycol (PEG).
Bottom: plastic (1 mm thick, polystyrene) or imaging bottom (0.18 mm thick, IBIDI polymer).
How do I choose between plastic or imaging bottom plates?
The choice of the plate bottom depends on your application needs. For high resolution imaging of immunostainings at 10X or higher magnification, imaging bottom (0.18 mm, IBIDI polymer) is preferred. For other types of assays, use plastic bottom.
What are the microwell sizes available?
Our Gri3D® plates come with various microwell sizes ranging from 100 μm to 1500 μm. Learn more about all the options by looking at our product catalogue.
- Standard microwell diameter (μm): 400 (121 μwells), 500 (73 μwells), 600 (55 μwells), 800 (31 μwells).
- Custom microwell diameter (μm): 100 (1677 μwells), 200 (511 μwells), 300 (211 μwells), 1000 (19 μwells), 1500 μm (7 μwells).
** the number of μwells per wells is given for a 96 wellplate.
To help you get started, we propose sample plates to test the different microwell sizes (only available for standard options). These special plates contain 1 column filled with hydrogel for each of the standard microwell sizes (400 – 800 μm). Learn more about the sample plates here.
How do I choose microwell size?
The purpose of having different diameters of microwells is to accommodate different organoid types and sizes. If your organoids are small or develop in a short time from low cell amounts, smaller microwell diameters will allow you to get a higher microwell density per area, therefore resulting in more datapoints per well. If you plan to grow your organoids to for longer times or to larger sizes, you can opt for larger microwell sizes.
We suggest that you start with our sample plates that contain 1 column filled with hydrogel for each of the standard microwell sizes (400 – 800 μm). This will enable you to determine the optimal microwell size for your application. If you can share the organoid model that you wish to grow, our applications team will provide you with the best microwell size range to test. You can send an email to [email protected].
Learn more about all the options by looking at our product catalogue. Learn more about the sample plates here.
What is the difference between standard and custom products?
Gri3D® standard plates come with microwell diameter sizes ranging from 400 to 800 μm. It can be ordered in 2 formats: 96 wells (full 96 wellplate) or 24 wells (in 96 wellplate). It is available directly on the webshop or by requesting a quote to [email protected] .
Gri3D® custom plates come with extended microwell diameter sizes (from 100 to 1500 μm). It can be ordered on 96 wellplate or 24 wellplate formats in a column-basis, from 1-column to a full plate. Only one microwell size can be chosen per plate. Production of custom products is upon demand with 5% extra fee and increased delivery time. The order is only possible by requesting a quote to [email protected].
GRI3D® PLATE USE
Where can I find protocols describing Gri3D® use?
Gri3D® plates are supplied with a protocol that details Gri3D® use. Scan the QR code on the plate and download it. You can also access all protocols here.
What cells have been tested in Gri3D®?
Our Gri3D® plates has been used to grow various models including but not limited to intestinal organoids, liver organoids, colorectal and pancreatic cancer organoids, blood-brain barrier organoids. Learn more about the various applications here.
How should I prepare Gri3D® for cell seeding?
First, aspirate the storage buffer from the wells, both in the pipetting port and in the cell seeding chamber. Add 150 μl (in Gri3D®-96) or 800 μl (in Gri3D®-24) of your medium of interest in the pipetting port and leave the plate for at least 15 minutes in the incubator or 30 minutes at room temperature. Next, remove medium from both the pipetting port and the cell seeding chamber and proceed with cell seeding.
How to aspirate off the media from the inner cell seeding ring without damaging the hydrogel?
With an aspirator and a Pasteur pipette, first remove the liquid from the pipetting port. Then, carefully access the cell seeding chamber and aspirate the remaining buffer until the microwell arrays become visible (full buffer removal is not necessary). For that, slide your pipette tip on the side of the well until you feel a resistance – the seeding ring; aspirate from there without touching the hydrogel.
How many cells do I need to seed in the microwells?
You need 50 ul (in Gri3D®-96) or 200 μl (in Gri3D®-24) of a single cell suspension per well. The cell density needs to be optimized for your application and depends on the growth rate of cells and desired final size of the organoid. You can control the organoid size by adjusting the starting cell seeding density. See details in the protocol provided with product or check our resources for more information on our established models.
How to evenly seed cells onto microwells?
Remove media until you see the hydrogel microwell pattern appear. Make sure your single cell suspension is well mixed and add 50 μl (in Gri3D®-96) or 200 μl (in Gri3D®-24) of cell suspension on top of the hydrogel microwells. Mix well between pipetting steps.
What is the typical seeding uniformity?
The seeding ring surrounding the hydrogel allows separation from the pipetting port and has a meniscus breaking effect which enables homogeneous cell seeding within a well. Variations in cell numbers between microwells in the same well are less than 5%.
Is it normal to see cells in the pipetting port after seeding?
If the seeding step is correct, there should not be any cells in the pipetting port after seeding. There are two potential causes for appearance of cells in the pipetting port: (1) the pipetting port wasn’t properly dried before seeding; (2) volume is above the seeding chamber volume (50 μl). Make sure the pipetting port is dry and that you add the correct seeding volume to prevent this effect.
How long do organoids take to form?
The formation time of the organoids strongly depends on the type of cells used, their proliferation rates and the cell seeding density used. These parameters need to be optimized for your application. It can take 2-5 days before the cells are fully compact.
How long can organoids be cultured in the microwells?
Organoids or spheroids can be cultured for as long as desired. For example, our embryonic stem cell-derived retinal organoids were cultured for 26 days, and we could culture primary human hepatocyte spheroids for 21 days. Check our resources for more information.
Can I establish co-cultures with multiple cell types on Gri3D®?
Yes. You can aggregate multiple cell types by mixing them before cell seeding, as we did for our blood-brain barrier organoid model (see application note). One can also add a second cell type such as immune cells for co-culture with the organoids once they are formed. Check for our T-cell killing assay application note for more information. Consult our protocols to learn how to do it.
How easy is it to change the medium?
Medium change is very simple. Our uniquely designed pipetting port, adjacent to the microwells, allows safe medium exchange or compound exposure without organoid loss. Simply remove 150 μl from the pipetting port and add the same volume of fresh medium.
How to prevent disruption of organoids already in the microwells during media exchange?
Our uniquely designed pipetting port, adjacent to the microwells, allows safe medium exchange or compound exposure without organoid loss. Simply remove 150 μl from the pipetting port and add the same volume of fresh medium.
How often do I change the medium?
Medium change frequency depends on the cell types and specific protocols. Medium can be changed every 2-3 days or as frequently as needed.
Do I need to add extracellular matrix (ECM) to my organoid cultures on Gri3D®? If so, how often and how much?
ECM needs depend on the organoid type. For organoids which are expanded embedded in basement membrane extract (BME), it may be necessary to mix ECM with the culture medium to allow for their 3D development and growth. In the case of Matrigel®, organoids usually need between 1.5 – 2% Matrigel®, to be added at every medium change. Consult our protocols to learn more or ask our experts at [email protected].
What types of ECM are compatible with Gri3D®?
Gri3D® is compatible with a range of ECM gels that can be diluted in media including but not limited to Collagen-I, Matrigel®, Matrigel® growth factor reduced BME, Cultrex BME, laminin.
Do I need to coat Gri3D® plates?
No, PEG is naturally cell-repellent, so no coating is needed before seeding the cells.
Do I need to centrifuge Gri3D® plates after cell seeding?
No, Gri3D® technology enables cell seeding and aggregation in a single pipetting step without centrifugation. The seeding ring surrounding the hydrogel allows separation from the pipetting port and has a meniscus breaking effect which enables homogeneous cell seeding within a well. Learn more about the technology here.
Do I need to use all wells at the same time?
Our recommendation is single-use of our Gri3D® plates. However, if one experiment didn’t use all wells in a plate, the remaining wells can be used in a subsequent experiment (within a few days), as long as the hydrogel microwells stay hydrated and that sterility is maintained. Before use, check if the unused microwells are still hydrated. Attention, this should not be done after PFA-fixation of wells.
Can I reuse Gri3D® after washing?
No, we do not recommend plate reuse after washing as we cannot guarantee the quality when reusing it.
How can I avoid damaging the hydrogel?
To avoid damaging the hydrogel, always use pipetting port for media removing and loading. When seeding the cells avoid reaching the hydrogel.
What happens if I reach the hydrogel?
Our PEG hydrogel is sensitive. If the hydrogel is disturbed by means of a pipet tip, it may damage the microwells, thus risking microtissue loss.
GRI3D® ASSAY
Once organoids are formed in each well, how shall we proceed to assays?
Once the organoids are formed, you can use them directly in your assay of choice. For example, if your assay is imaging, you image the cells directly on the plate after incubation with your dye of interest or proceed to your optimized immunostaining protocol. You can also use the supernatant for your assays of interest or recover single organoids from the well for other downstream analyses. Learn more about the compatible assays by checking our resources.
Can I do immunostaining assays on Gri3D®?
Yes, all steps of immunofluorescence can be performed on Gri3D®, from fixation, permeabilization, blocking, antibody incubation all the way to imaging. We recommend the use of imaging-bottom plates for optimal imaging results.
Can I image my samples on Gri3D®?
Gri3D® is compatible with regular transmitted light and fluorescence imaging, also in confocal mode. On Gri3D®, growing organoids are positioned in confined areas (the microwells) and in a single focal plane, allowing the imaging of multiple organoids at the same time. This allows the establishment of efficient image-based workflows which minimize the acquisition time and maximise the data extracted.
Can I use Gri3D® for organoid screening?
Gri3D® comes in SLAS-standard 96-wellplate format and is well suited for screening programs as it generates a high volume of organoids per well, allowing multiple replicates per well to be generated and thus maximizing the datapoints per well. Moreover, as there are multiple organoids in a single well, this allows for multiple assays to be performed independently of their limit of detection.
Is the plate compatible with fluorescent / colorimetric (O.D.) readings?
Fluorescence and colorimetric based assays can be performed on Gri3D®. However, Gri3D® is transparent so we recommend sampling out in an assay plate (white-well or black-well plate) for optimal assay results.
How can I assess organoid viability on Gri3D®?
Gri3D® is compatible with various cell viability assays including fluorescence image-based assays such as LIVE/DEADTM Kit or luminescence-based assays like CellTiter-Glo® 3D.
Does the hydrogel interfere with CellTiter-Glo® 3D or other luminescence-based assays?
No, CellTiter-Glo® 3D reagents contain a lytic compartment that lyses cells and a compound that reacts to the presence of ATP coming from the lysed cells. None of these have an effect on PEG, which is a cell-inert compound. Thus there is no interference of the hydrogel with luminescence- based assays.
Can I sample out media from Gri3D®?
Yes, you can safely sample media out by using the pipetting port for downstream assays such as mass spectrometry MS.
How can I harvest the organoids?
As cells grow in an open and solid matrix-free environment, they can be easily harvested from the microwells with a simple pipetting step on top of the growing organoids – no need for cumbersome extraction protocols. Single organoids or homogeneous organoid populations can then be processed for passaging or downstream analyses. See protocol.
Is it okay that the harvested organoids/spheroids contain some of the hydrogel?
Yes, PEG is cell-inert and can be removed after organoid centrifugation.
Can I extract RNA/DNA/protein from my 3D cultures on Gri3D®?
After harvesting the organoids, you can proceed with your in-house protocol to extract DNA, RNA or protein. Depending on the amount of microtissues and their size, you may have to pool several wells to have enough starting material.
Can I pick single organoids from the microwells?
Yes, as cells grow in an open and solid matrix-free environment, one can pick single organoids from the microwells.
Can I cryosection my organoid samples?
Yes, samples in the microwells can be fixed and cryopreserved by successive incubations in sucrose gradients. Then, the hydrogel containing the organoids can be scooped out, embedded in OCT, snap-frozen and cryosectioned (see publication).
How does organoid culture on Gri3D® compare to other organoid culture methods?
1. Homogeneity and reproducibility – as organoids grow independently from a single cell suspension in each of the microcavities, this results in homogenously sized and positioned organoids when compared to solid-ECM embedding (see poster). Furthermore, all organoids are exposed to the same concentrations of growth factors, cytokines or compounds that are in the media, without any gradient, thus further increasing reproducibility.
2. Throughput – as you are no longer limited to a single replica per well, but up to hundreds of microtissues per well (depending on the microwell size), thus increasing the throughput and decreasing reagents consumption.
3. Safe medium exchange – thanks to the pipetting port, no microtissue loss occurs. The cell compartment is only accessed for seeding, and media changes are performed in an independent inlet, the pipetting port, without disturbing the cells during spheroid formation and avoiding microtissue loss during culture.
4. Imaging compatibility – from simple brightfield imaging to complex immunostainings in confocal mode directly on Gri3D®. No diffraction and no need to harvest the cells.
5. Automation friendly – SBS/ANSI format of the plate allows full automation of cell culture process in the seeding chamber as well as the pipetting port.
6. Simple assay performance and high sensitivity – with low volumes, and without losing microtissues. One can incubate the reagents by using the pipetting port. As multiple microtissues grow in a single well on Gri3D®, assay sensitivity is increased, so these can be performed independently of their limit of detection.
7. Customisation – in Gri3D®, it is possible to culture differently sized microtissues by choosing different microwell sizes and seeding densities.
What are the application areas where Gri3D® has been used?
To date, Gri3D® has been used to establish robust and reproducible organoid cultures of multiple organs, such as intestine, liver or pancreas, from both healthy as well as tumor origin. Moreover, it has been used to establish co-cultures with immune cells to look at their tumor killing capacity. (Dutta et al., 2021). Gri3D® has also been used to form different spheroid types, including but not limited to cancer cell line spheroids (HCT116) (Brandenberg et al, 2020), primary human hepatocyte spheroids, blood-brain barrier spheroids (a self-assembling culture of astrocytes, pericytes and endothelial cells) (Simonneau et al, 2021), bone marrow spheroids (with mesenchymal stem cells and HUVECs) (Giger et al, 2022).
Check our resources to learn more.
In terms of applications, Gri3D® is used in ADME-Tox assays (efficacy testing of drugs, safety profiling), basic and applied research in neuroscience, oncology, immunology and developmental biology, and for cell therapy purposes.
