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Frequently Asked Questions
Glass Bottom Culture Dishes
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
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Dish Utilization
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| Q-DU1 |
How
are glass bottom dishes typically used?
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| Q-DU2 |
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?
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| Q-DU3 |
How
can I search for published methods utilizing MatTek's glass bottom dishes?
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| Q-DU4 |
What
should I do if my cells won't grow?
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| Q-DU5 |
Can
the coverslip be removed from the glass bottom dish?
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| Q-DU6 |
How can I control
the temperature of the glass bottom dishes for in situ (live) microscopy?
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| Q-DU7 |
Are
the glass bottom dishes good for fluorescence microscopy?
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| Q-DU8 |
How
can I use the glass bottom dishes for Nomarski Differential Interference Contrast (DIC) microscopy?
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| Q-DU9 |
Why
would one want to use glass bottom dishes containing gridded coverslips? What is the size of
the grid?
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Q-DU10
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Why
are the 50 mm glass bottom dishes useful for microinjection and maintaining a constant atmosphere
in the dish?
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Dish Properties
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| Q-DP1 |
What
is the adhesive used to attach the coverslips to the petri dishes?
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| Q-DP2 |
How
long can the dishes be stored?
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| Q-DP3 |
Over
what temperature range can the glass bottom dishes be used?
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| Q-DP4 |
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?
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| Q-DP5 |
How
deep is the well of the glass bottom dish?
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| Q-DP6 |
What
is the chemical compatibility of Glass Bottom Dishes and Multi-Well Plates? Can they be used
with organic solvents?
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| Q-DP7 |
Why
would one want to use glass bottom dishes containing gridded coverslips? What is the size of
the grid?
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Coatings
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| Q-DC1 |
What
is the molecular weight range of the poly-d-lysine used to coat the PDL coated glass bottom
dishes?
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| Q-DC2 |
Why
are the dishes coated with poly-d-lysine (instead of poly-l-lysine)?
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| Q-DC3 |
What
type of collagen is applied to the collagen coated glass bottom dishes (part #: P35GCol-x-xx-C
or P50GCol-x-xx-F)?
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| Q-DC4 |
Are
the poly-lysine or collagen coatings going to affect the optical properties of the glass bottom
dishes?
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Special Orders
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| Q-SO1 |
Other
than the standard 10mm and 14mm hole sizes, do the glass bottom dishes come with any alternative
hole diameters?
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| Q-SO2 |
How
long will it be before my standard order or special order glass bottom dishes will ship?
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How are glass bottom dishes typically used?
MatTek's
glass bottom dishes come uncoated or coated with poly-d-Lysine or collagen. All dishes are gamma
irradiated so all dishes must be handled in a sterile environment to prevent contamination.
A general procedure for their use follows.
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.
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| A-DU02
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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.
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A-DU03
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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.)
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A-DU04
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What
should I do if my cells won't grow?
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 dishes themselves. They purchase the uncoated dishes
and use the HCl pretreatment (see below) along with their coating of choice. To try a sample
of non-coated glass bottom dishes, please give our customer service department a call (800-634-9018).
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 H20.
3. Apply your coating to the dishes. |
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A-DP1
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What
is the adhesive used to attach the coverslips to the petri dishes? |
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Although the
specific identity of the adhesive is proprietary, the adhesive used is a non-toxic silicone
that has been shown to be compatible with a broad variety of cells including primary neurons
and many other fastidious cells. |
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A-DU05
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Can
the coverslip be removed from the glass bottom dish?
Yes. However, for most applications, cells
growing in the glass bottom dish can be viewed without removal of the coverslip. However, if
necessary (e.g. for long term storage purposes), the coverslip can be removed using the following
procedure:
1. Order Part # PDCF OS 30 (Dow Corning fluid OS 30, Price = $20/50 ml available from MatTek)
2. Invert the cover of the dish.
3. Pipette 1.0 ml of OS 30 into the inverted cover.
4. Place the bottom of the dish in the cover. Make sure that the liquid is touching the bottom
of the coverslip.
5. Allow the dish to sit in the OS 30 for 45 minutes at room temperature.
6. Dry the bottom of the coverslip with an absorbent paper towel.
7. Place the dish on a clean surface. Using forceps, press down on the edge of the coverslip
to separate the coverslip from the dish.
Note: If the above procedure is followed, the PDCF OS 30 liquid
will not contact the cells and will
not disrupt cells on the coverslip or the staining thereof. |
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A-DC1
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What
is the molecular weight range of the poly-d-lysine used to coat the PDL coated glass bottom
dishes?
The poly-d-lysine used to coat the
P35GC-x-xx-C or P50GC-x-xx-F glass bottom dishes is in the molecular weight range of 70,000-150,000
Daltons. |
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A-DC2
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Why
are the dishes coated with poly-d-lysine (instead of poly-l-lysine)?
Both untreated glass and cells are negatively
charged. Poly-lysine is applied to the glass surface to make it positively charged, thereby
increasing electrostatic attraction between the glass surface and the cells and thus favoring
cell attachment. Poly-d-lysine is favored because the d-enantiomer is less prone to protease-mediated
breakdown than the naturally-occurring l-enantiomer. Otherwise, Poly-d-Lysine and Poly-l-Lysine
are equivalent. |
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A-DC3
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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.
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A-DC4
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Are
the poly-lysine or collagen coatings going to affect the optical properties of the glass bottom
dishes?
The coatings are monolayer coatings which
do not affect the optical properties of the glass bottom dishes. |
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A-DP2
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How
long can the dishes be stored?
Uncoated dishes have no maximum shelf life.
Coated dishes can be stored in the dark for up to 2 years. |
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A-DP3
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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.
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A-DU6
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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
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A-DP4
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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:
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Coverslip No.
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Thickness (mm)
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0
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0.085-0.13
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1.0
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0.13-0.16
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1.5
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0.16-0.19
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2.0
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0.19-0.23
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A-DU7
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Are
the glass bottom dishes good for fluorescence microscopy?
Yes. The glass bottom dishes are excellent
for fluorescent microscopy. Important glass properties are:
1. Incident ultraviolet rays with wavelengths longer than 320 nm do not cause fluorescence.
2. Mercury lines at 334 and 365 nm do not create auto-fluorescence. (Note: For mercury illumination,
filter out the mercury lines with wavelengths shorter than 313 nm to obtain best possible results.)
3. Refractive index (@ 20°C): nd= 1.5230 tolerance ± 0.0015
4. Abbe number V = 55.
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A-DU8
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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.
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A-DP5
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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.
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A-DU9
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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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A-SO1
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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).
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A-SO2
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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.
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A-DU10
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Why are the 50 mm glass bottom dishes useful
for microinjection and maintaining a constant atmosphere in the dish?
The 50 mm glass bottom
dishes (part #'s beginning with P50G) are useful for:
a) Microinjection: The larger diameter (50 mm) and the lower side wall (7 mm) allows easier
access to cells in microinjection experiments.
b) Atmosphere maintenance: The 50 mm dish has a cover that snaps onto the dish bottom and thereby
prevents loss of the 5% CO2 atmosphere while the dish is out of the incubator. This can be important
for experiments in which dishes will be observed for extended periods.
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A-DP6
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What is the chemical compatibility of the Glass
Bottom Dishes and Multi-Well Plates? Can they be used with organic solvents?
The
body of the glass bottom dishes and multi-well plates is made from polystyrene. Therefore, they
have limited compatibility with organic solvents. Please see the chemical compatibility table.
| Solvent |
Chemical Compatibility |
| Acetone |
Poor |
| Ammonium hydroxide (1N) |
Fair |
| Ammonium hydroxide (25%) |
Fair |
| Aniline |
Good |
| Butanol |
Good |
| Chloroform |
Poor |
| Dimethylformamide |
Poor |
| Dimethylsulfoxide(DMSO) |
Poor |
| DMSO/H2O (20/80)
| Good |
| Dioxane |
Poor |
| Ethanol |
Good |
| Hexane |
Poor |
| Hydrochloric acid (25%) |
Good |
| Hydrochloric acid (concentrated) |
Fair |
| Methanol |
Good |
| Methyl ethyl diketone |
Poor |
| Methylene chloride |
Poor |
| Nitric acid (25%) |
Poor |
| Nitric acid (concentrated) |
Poor |
| Sodium hydroxide |
Good |
| Toluene |
Poor |
| Xylene |
Poor |
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