ibidi Glass Bottom Dish User manual

Instructions Glass Bottom Dish 35 mm
The ibidi product family is comprised of a variety of µ–Slides and µ–Dishes, which have all been
designed for high–end microscopic analysis of fixed or living cells.
The glass bottom versions of the µ–Slides and µ–Dishes are especially designed for TIRF, super reso-
lution and single molecule applications.
The Glass Bottom Dish 35 mm is a standard glass bottom dish for cell culture and high resolution
microscopy.
Material
The Glass Bottom Dish 35 mm is made of a standard 35 mm
Dish, but with a glass coverslip bottom. It is not possible to
detach the bottom. The Dishes are not autoclavable since
they are temperature stable only up to 80°C / 175°F.
Optical Properties Glass Bottom Dish 35 mm
Refractive index nD1.523
Abbe number 55
Thickness No. 1.5
(170 µm (+20 µm/-10 µm))
Material Schott borosilicate glass,
D 263M
Attention!
Be cautious when handling ibidi labware products
with glass bottom! The glass coverslip or glass slide
is very fragile and might break easily. Handle with
care to avoid physical injury and damage to devices
through leakage of the medium.
Geometry
Geometry of the Glass Bottom Dish 35 mm
Diameter dish 35 mm
Volume 2 ml
Growth area 3.14 cm2
Coating area using 400 µl 3.7 cm2
Diameter observation area 20 mm
Height with / without lid 11.7 mm/9.9 mm
Bottom Glass coverslip No. 1.5
Surface and Coating
The Glass Bottom Dish 35 mm is manufactured with an un-
coated glass coverslip. Washing steps (e.g. with PBS) be-
fore cell seeding can remove glass dust which is advanta-
geous for direct cell growth on the surface.
Protein coatings increase direct cell growth of adherent
cells. Specific coatings on glass are possible following this
protocol:
• Prepare your coating solution according to the man-
ufacturer’s specifications or reference. Prepare your
Glass Bottom Dish 35 mm. Adjust the concentration
to a coating area of 3.7 cm2and a coating volume of
400 µl.
• Apply 400 µl into the growth area. Make sure that
the entire bottom is covered with liquid easily tilting
or shaking the µ–Dish. Put on the lid and leave at
room temperature for at least 30 minutes.
• Aspirate the solution and wash. Optionally, let dry
at room temperature.
Detailed information about coatings is provided in Appli-
cation Note 08 ”Cell culture coating”.
Shipping and Storage
The µ–Slides, µ–Dishes and µ–Plates are sterilized and
welded in a gas-permeable packaging. The shelf life under
proper storage conditions (in a dry place, no direct sun-
light) is listed in the following table.
Conditions
Shipping conditions Ambient
Storage conditions RT (15-25°C)
Shelf Life
Glass Bottom 36 months
Glass Bottom Dish 35 mm Page 1 Version 1.1 (2018-07-11)

Instructions Glass Bottom Dish 35 mm
Seeding Cells
Depending on your cell type, application of a 4–9 ×
104cells/ml suspension should result in a confluent layer
within 2–3 days.
• Trypsinize and count cells as usual. Dilute the cell
suspension to the desired concentration.
• Apply 400 µl cell suspension into the inner well of
the µ–Dish. Avoid shaking as this will result in inho-
mogeneous distribution of the cells.
• After cell attachment add additionally 1.6 ml of pure
medium to ensure optimal grow conditions.
• Cover the µ–Dish with the supplied lid. Incubate at
37°C and 5 % CO2as usual.
We recommend not to fill more than the indicated to-
tal volume into the Glass Bottom Dish 35 mm in order to
avoid the liquid contacting the lid.
Undemanding cells can be left in their seeding medium for
several days and grow to confluence there. However, best
results are achieved when the medium is changed every
2–3 days. Carefully aspirate the old medium and replace
it by up to 2 ml fresh medium.
Glass Bottom Dish Selection Guide
Glass Bottom
Dish 35 mm
µ–Dish 35mm, high
Glass Bottom
Lid normal with Lid Lock po-
sition
Evaporation medium low
Coverslip No. 1.5 No. 1.5H
Thickness 170 µm
(+20 µm/-10 µm)
170 µm (±5 µm)
Packaging 10 pieces 1 piece
Note:
If cultivated in a dry atmosphere, evaporation can
harm the cells during longer experiments. Exchange
the medium every 1-2 days to minimize evaporation-
mediated osmolality shifts.
Optional Glass Coverslip Cleaning Protocol
The Glass Bottom Dish 35 mm is made with an uncoated
glass coverslip. For improved cell attachment, glass sur-
face modifications and other applications, the glass cov-
erslip of the Glass Bottom Dish 35 mm can be cleaned by
following the protocol below.
• Remove lids and immerse products in ddH2O in an
appropriately sized beaker.
• Sonicate for 10 minutes.
• Decant the ddH2O completely.
• Add 1 M HCl.
• Sonicate for 10 minutes.
• Decant the HCl completely and wash twice with
ddH2O. Decant the ddH2O completely.
• Add 2-propanol (absolute).
• Sonicate for 10 minutes.
• Aspirate the 2-propanol completely. Make sure that
all products are completely dry. Wash twice with
ddH2O and aspirate the ddH2O completely.
• Add ethanol (absolute).
• Sonicate for 10 minutes.
• Aspirate the ethanol completely. Make sure that
all products are completely dry. Wash twice with
ddH2O.
• Sonicate in ddH2O for 10 min.
• Decant ddH2O and blow dry carefully with canned
air or clean nitrogen gas.
Modifications of this protocol including acids, bases, alco-
hols and detergents are possible. Please check the chem-
ical compatibility list on www.ibidi.com for compatibil-
ity. Make sure to handle the glass-bottomed products with
care. The glass coverslips may break during mechani-
cal handling. For best results, use a custom-made Teflon
holder.
Glass Bottom Dish 35 mm Page 2 Version 1.1 (2018-07-11)

Instructions Glass Bottom Dish 35 mm
Cell Microscopy and Solvents for Fixation
To analyze your cells, no special preparations are neces-
sary. Cells can be observed live, or fixed directly in the
Glass Bottom Dish 35 mm, preferably on an inverted micro-
scope. Due to the thin bottom, high resolution microscopy
is possible. The material is compatible to most fixatives,
like acidic acid, alcohols and PFA. For a full list of compat-
ible solvents and more information on chemical compati-
bility, please visit the FAQ section on www.ibidi.com. For
optimal results in fluorescence microscopy and storage of
stained probes ibidi provides a mounting medium (50001)
optimized for µ-Dishes and µ-Slides.
Ordering Information
µ–Dish 35 mm, high
Cat. No. Description
81156 µ–Dish 35 mm, high ibiTreat:  35 mm, high wall (2 ml volume), #1.5 polymer coverslip, tis-
sue culture treated, sterilized
81156-400 µ–Dish 35 mm, high ibiTreat, Bulk Pack:  35 mm, high wall (2 ml volume), #1.5 polymer
coverslip, tissue culture treated, sterilized
81151 µ–Dish 35 mm, high Uncoated:  35 mm, high wall (2 ml volume), #1.5 polymer coverslip,
hydrophobic, sterilized
µ–Dish 35 mm, high Grid–500
Cat. No. Description
81166 µ–Dish 35 mm, high Grid-500 ibiTreat :  35 mm, high wall (2 ml volume), grid repeat distance
500 µm, #1.5 polymer coverslip, tissue culture treated, sterilized
81161 µ–Dish 35 mm, high Grid-500 Uncoated:  35 mm, high wall (2 ml volume), grid repeat dis-
tance 500 µm,#1.5 polymer coverslip, hydrophobic, sterilized
Glass Bottom Dish 35 mm
Cat. No. Description
81218-200 Glass Bottom Dish 35 mm:  35 mm, high wall (2 ml volume), #1.5 (170 µm (+20 µm/-10 µm))
D 263 M Schott glass, sterilized, 200 pieces
81218-800 Glass Bottom Dish 35 mm:  35 mm, high wall (2 ml volume), #1.5 (170 µm (+20 µm/-10 µm))
D 263 M Schott glass, sterilized, 800 pieces
µ–Dish 35 mm, high Glass Bottom
Cat. No. Description
81158 µ–Dish 35 mm, high Glass Bottom:  35 mm, high wall (2 ml volume), #1.5H (170 ±5 µm) D
263 M Schott glass, sterilized
81158-400 µ–Dish 35 mm, high Glass Bottom, Bulk Pack:  35 mm, high wall (2 ml volume), #1.5H (170
±5 µm) D 263 M Schott glass, sterilized
Glass Bottom Dish 35 mm Page 3 Version 1.1 (2018-07-11)

Instructions Glass Bottom Dish 35 mm
µ–Dish 35 mm, low
Cat. No. Description
80136 µ–Dish 35 mm, low ibiTreat:  35 mm, low wall (800 µl volume), #1.5 polymer coverslip, tissue
culture treated, sterilized
80131 µ–Dish 35 mm, low Uncoated:  35 mm, low wall (800 µl volume), #1.5 polymer coverslip,
hydrophobic, sterilized
For research use only!
Further technical specifications can be found at www.ibidi.com. For questions and suggestions please contact us by e-mail [email protected]
or by telephone [phone]. All products are developed and produced in Germany.
© ibidi GmbH, Am Klopferspitz 19, 82152 Martinsried, Germany.
Glass Bottom Dish 35 mm Page 4 Version 1.1 (2018-07-11)