Click on a subject heading below to view questions and answers relating to your selection. Links throughout the answers will guide you to further information on our website or from other sources. Should you have any further questions, please contact MatTek Corporation at DishInfo

How can I control the temperature of the glass bottom dishes for in situ (live) microscopy?

1. Maintain sterility: Open dishes in a sterile environment (e.g. laminar flow hood).

2. Pre-equilibrate dishes: Incubate the dishes with culture medium. Pipet 2-3 ml of medium into the 35 mm dishes or 3-4 ml into the 50 mm dishes and incubate at 37° C for 15 minutes.

3. Add cell suspension to microwell: Remove the culture medium by aspiration and plate cells onto the glass surface. Pipet 250µl of the cell suspension (cells suspended in culture medium) into the 10 mm diameter microwells or 500µl of cell suspension into the 14-mm microwells. Incubate the dishes for 1 hour at 37° C.

4. Add additional medium: After 1 hour, gently fill the remainder of the dish with medium Add 2-3 ml to the 35 mm dishes or 3-4 ml for the 50 mm dishes. Note: After the initial one hour period to allow cells to attach to the glass surface, it is important to fill the dish to normal levels in order to minimize the effects of evaporation and to avoid inducing changes in osmolarity.

What type of glass bottom dish should I use to grow my cells? How can I use the glass bottom dishes to look at tissue slices?

It is hard to predict which type of glass bottom dishes (uncoated, poly-d-lysine coated, or collagen coated) will work best with your specific cell type. Many transformed or cancerous cell lines will grow on uncoated dishes. Poly-lysine coated dishes work well for neuronal culture and for many primary cells; other cells prefer a collagen coating. Also, many researchers purchase our uncoated dishes and apply their own specialized coating. You can also Google search for published methods utilizing glass bottom dishes and your cells (see Q#3).

The glass bottom dishes can also be used to image tissue slices. The slices are adhered to the dishes using Cell-Tak (BD BioSciences see: http://www.bdbiosciences.com/discovery_labware). A research paper utilizing MatTek's glass bottom dishes to perform confocal microscopy on brain slices is available through this link.

How can I search for published methods utilizing MatTek's glass bottom dishes?

You can search Google for publications by using search terms: 'MatTek, glass bottom dishes, and your cell type". For instance, the search terms 'MatTek', 'glass bottom dishes' and 'Hela' will yield numerous articles utilizing MatTek's glass bottom dishes to culture Hela cells. You can also search by type of microscopy. Use the terms: "MatTek, glass bottom dishes, and your type of microscopy." Note: If the exact glass bottom dish part number is unspecified in a literature article, it is safe to assume that uncoated dishes were used (e.g., P35G-0-14-C, P35-1.5-14-C,
etc.)

What should I do if my cells won't grow or to improve growth in the glass bottom dishes?

A: Although our poly-d-lysine or collagen coating works well for the vast majority of customers, some customers find it necessary to coat the glass bottom dishes themselves. They purchase the uncoated dishes and use the following HCl pretreatment along with their coating of choice.

1. Under sterile conditions, pipette 250µl of 1 N HCl onto non-coated 10-mm glass bottom dishes (P35G-x-10-C or P50G-x-10-F); or pipette 500µl of 1 N HCl into the 14-mm glass bottom culture dish (P35G-x-14-C or P50G-x-14-F).

2. After 15 minutes, decant the HCl and rinse the dish 3x with phosphate buffered saline (PBS) and 2x with ultrapure H2O.

3. Apply your coating to the dishes.

4. Add a similar volume of the medium in which you will plate your cells to pre-equilibrate the glass surface. Incubate the medium in glass bottom dishes for 15 mins at 37 C. Remove the medium and then plate your cells.

To try a sample of non-coated glass bottom dishes, complete the Free Sample request Form.

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What type of collagen is applied to the collagen coated glass bottom dishes (part #: P35GCol-x-xx-C or P50GCol-x-xx-F)?

The collagen used to coat the P35GCol-x-xx-C or P50GCol-x-xx-F glass bottom dishes is type 1 rat tail collagen.

Over what temperature range can the glass bottom dishes be used?

The glass bottom dishes can be cooled to -196ºC (temperature of liquid nitrogen). The dishes will become disfigured at 55ºC (131ºF) thus temperatures above 50ºC are not recommended. This means that the dishes CANNOT be autoclaved.

How can I control the temperature of the glass bottom dishes for in situ (live) microscopy?

In order to approximate physiological conditions, the temperature of the medium contained within the glass bottom dishes can be controlled by using a microscope stage heater and an appropriate stage adapter.

For use with the P35G dishes (Corning 35 mm dishes) only: Culture dish heaters (part # DH-35), microscope stage adapters (part #: SA-microscope type), heater controller (part # TC324-B), and connecter cable (part # CC-28) are available from Warner Instrument Corporation. Information is available on line at: http://www.warneronline.com/products.cfm

What are properties of the glass used in the glass bottom dishes? What are the thicknesses of the different coverslips used in the glass bottom dishes?

MatTek uses the highest quality, borosilicate German glass coverslips in its glass bottom dishes. The coverslip properties are as follows:

1. Highest hydrolytic resistance (hydrolytic class 1)
2. Excellent resistance to chemicals
3. Emission of alkali approximately 15 to 24 µg Na2O/g glass

The thickness of the glass coverslips depends on the Coverslip No., as follows:

How can I use the glass bottom dishes for Nomarski Differential Interference Contrast (DIC) microscopy?

You will need to order both a glass bottom dish and a glass cover. Order any P35G dish along with a glass cover (Part#P35-GTOP-0-20-C). The glass covers can be re-used following resterilization of the covers by soaking them in 70% ethanol for 30 minutes.

How deep is the well of the glass bottom dish?

The depth of the wells in the glass bottom dishes depends on the type of dish as follows:

1. Corning 35mm: 0.70 - 0.75mm
2. Falcon 50mm: 1.00-1-10mm
3. Falcon 35mm: 1.15 - 1.25mm*
4. Nunc 35mm: 0.80 - 0.85mm*
5. Falcon 6-well plate: 1.45-1.55 mm
6. Falcon 12-well plate: 1.45-1.55 mm
7. Falcon 24-well plate: 1.10-1.20 mm
8. Falcon 96-well plate: 1.05-1.25 mm
*Note: Glass bottom dishes made from dish types 3 and 4 are special order items.

Why would one want to use glass bottom dishes containing gridded coverslips? What is the size of the grid?

The gridded coverslips allow one to refer to specific cells and follow them over time. For instance, individual cells can be microinjected, returned to the incubator, and observed at multiple time points since each cell can be identified with a unique alpha-numeric coordinate in the dish. Glass bottom dishes containing gridded Bellco Glass coverslips are available. Part numbers for standard gridded dishes are: P35G-2-14-C-GRID and P50G-2-14-F-GRID.

Grid size: The grid on P35G-2-14-C-GRID and P50G-2-14-F-GRID consists of 520 unique alphanumeric squares. Each square measures 600 microns x 600 microns. The line thickness is 20 microns.

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Other than the standard 10 mm and 14 mm hole sizes, do the glass bottom dishes come with any alternative hole diameters?

Glass bottom dishes with 20 mm holes are also standard products (e.g. Part # P35G-1.5-20-C or P35G-1.0-20-C). In addition, dishes with 5, 7, and 30 mm diameter holes are available on a special order basis. The 5 and 7 mm holes are useful when very expensive reagents need to be conserved. The 30 mm holes maximize the surface area for cell growth. Note: The 30 mm hole is available in the 50 mm dish only (e.g. Part #P50G-1.5-30-F or P50G-0-30-F).

How long will it be before my standard order or special order glass bottom dishes will ship?

Standard glass bottom dishes are typically available to ship on the day that a purchase order is received at MatTek.

For special order or custom size dishes, 4-8 weeks is a "worst case" lead time. The lead time mainly depends on sterilization which is performed off site on very large lots of dishes using gamma irradiation. Depending on when the special order is placed in relation to the next sterilization batch, the lead time can be shorter. In addition, the lead time can be shortened to 2-4 weeks if the initial order can be shipped non-sterile. In this case, the customer will need to sterilize the dishes for 30 minutes under the UV light in a cell culture hood.

For more accurate lead times for specific orders, please ask our customer service representatives.