Series KIT-3522        /     500 units per case.

1 case of the transparent 'WillCo-dish®KIT', contains:

500 Dishes & Lids,
500 Coverslips of 'German Glass D-263M' of which you decide, which of our types of glass you prefer.
500 3M 'Double Sided Adhesive (DSA) rings',
1 Tweezer and
1 Pair of Latex® gloves.

Assembly Device
To assemble the WillCo-dish®KIT 'Do-It-Yourself' dishes "Easy, Accurate/precise, Quick and Safe", it is necessary to
have the ASSEMBLY DEVICE, type: DEV-3522 (Comes separately).
The blue part of the Device is needed to position, center and bond the adhesive ring, to the bottom of the dish.
The red part of the Assembly Device, is needed to center - and bond the glass coverslip, to the bottom of the dish.

Other Packaging: 

Other options for delivery of your WillCo-dish®-es are in:

BLISTER-PACK   :      PET-G BLUE MED blister with Tyvek® cover;  'Single unit' packed.
                                          4 Dishes per blister x 30 blisters = 120 Units per case.
                                          STERILE R by Gamma irradiation, 10 kGy minimum.
                                          NOTE:  Products are sterile, until packaging is opened or damaged.

POUCH-PACK    :        PA/PE and Tyvek® (DuPont) sleeve.  'Multi unit' packed;
                                          20 Dishes per sleeve x 10 sleeves = 200 units per case.
                                          STERILE R, by Gamma irradiation, 10 kGy minimum.
                                          NOTE:  Products are sterile, until packaging is opened or damaged.

If you have just any questions concerning our products or packaging, please do not hesitate to contact us.

Your own shipper

In case you take care of your own transport, your shipper may want
to have the following information, on your goods.

SIZES, VOLUMES & WEIGHTS:

GWST-Series & GWSB-Series cases (Blister-Pack)
1 Case:  Size: 36 x 28 x 19 cm. / Volume: 0.02 m3 / Weight:  1.95 kg.
2 Case:  Size: 36 x 28 x 39 cm. / Volume: 0.04 m3 / Weight:  3.90 kg.
3 Case:  Size: 36 x 28 x 59 cm. / Volume: 0.06 m3 / Weight:  5.90 kg.
4 Case:  Size: 36 x 28 x 66 cm. / Volume: 0.08 m3 / Weight:  7.90 kg.
5 Case:  Size: 36 x 56 x 59 cm. / Volume: 0.12 m3 / Weight:  8.85 kg. (We add one empty case, for the right 3+3 shape).

HBST-Series & HBSB-Series cases (Pouch-Pack)
1 Case:  Size: 26 x 22 x 18 cm. / Volume: 0.01 m3   / Weight:  1.60 kg.
2 Case:  Size: 26 x 22 x 36 cm. / Volume: 0.02 m3   / Weight:  3.30 kg.
3 Case:  Size: 26 x 22 x 54 cm. / Volume: 0.03 m3   / Weight:  4.90 kg.
4 Case:  Size: 43 x 35 x 25 cm. / Volume: 0.037 m3 / Weight:  6.70 kg.
5 Case:  Size: 53 x 43 x 25 cm. / Volume: 0.057 m3 / Weight:  8.95 kg. (We add one empty case, for the right 3+3 shape).

WillCo-dish®KIT-Series cases (KIT-Pack)
1 Case:  Size: 36 x 28 x 19 cm. / Volume: 0.02 m3 /  Weight:  2.45 kg.
2 Case:  Size: 36 x 28 x 39 cm. / Volume: 0.04 m3 /  Weight:  4.90 kg.
3 Case:  Size: 36 x 28 x 59 cm. / Volume: 0.06 m3 /  Weight:  5.90 kg.
4 Case:  Size: 36 x 28 x 65 cm. / Volume: 0.08 m3 /  Weight:  9.90 kg.

Your WillCo Wells B.V. 'Customer Service-Team'

WillCo-dish®KIT 'Series KIT-3522'

Dear Colleague,

For these WillCo-dish®KIT 'Do-It-Yourself' Glass Bottom dishes, the 'Series KIT-3522', we use diameter
30 mm. glass, manufactured by SCHOTT. Supplier is Thermo Fisher (Former: Gerhard Menzel GmbH.).
For more specifications of the borosilicate D-263 M glass, you may want to visit their web site, to find all
applicable information on the 'glass bottom' of our WillCo-dish®'es.

Diameter:  30 mm.
Thicknesses we offer:

Class #1.5H:  170 micron +/- 5 micron, with flatness  (Ra) 0.005 mm. (H: Specially selected glass).

Class #1.5   :   Everage of 170 micron (160 - 190 micron), with (Ra) 0.01 mm., as well as (Ra) 0.005mm.
NOTE          :   We developed the KIT-Series, to be able to use any type of dish-bottom; coverglass, quartz,
                        artificial quartz, any type of plastic workable for you.

and:

Class #1      :   Everage of 140 micron (130 - 160 micron).
                        Flatness (Ra) 0.01 mm., only.
NOTE          :   We receive requests for these specificatios, not often, but we have plenty of #1 coverslips
                        for you in stock.

Your WillCo Wells B.V. 'Customer Service-Team'

WillCo-dish®KIT 'Series KIT-3522'

Flatness:

Speaking about Glass Bottom Dishes, there are two definitions of 'flatness'.

First, there is the flatness of the glass coverslip bottom of the dish, where the glass coverslip is bonded to the dish as such, that it is absolutely horizontal
on the warming stage (- and should be flush with the warming stage; there is no air buffer!).

Glass coverslip horizontal:
Positioned flat or horizontal means, that the light of the microscope passes through the glass bottom orthogonally (very important) and that your cells stay in
focus, when you move the warming stage to another position (E.g. but not limited too: Time Laps Microscopy).

Glass coverslip 'flush', with the stage:
When the glass coverslip is flush with the stage, it means that there is no air-buffer, between the glass and the warming stage. The WillCo-dish® design,
was - and is, the first dish to have this exceptional design. That is what makes the WillCo-dish®DESIGN of such a unique nature.
This feature too is very important, because it ensures an even distribution of the heat, from the warming stage to the media/liquid, inside the dish. All cells
will be evenly heated, which is essential for the well being of your precious cells and for the success of your work!

Note: If this is not the case, if there is an air-buffer between the warming stage and the bottom of the glass coverslip, an uneven distribution of the heat to your
cells, this will shorten their life dramatically and influence the results of your work.

Coverslip roughness:
Secondly, when discussing 'flatness' of the glass coverslip, we mean the surface flatness (Ra) or roughness of the surface of the glass.
We offer two specifications: Flatness Ra 0.01 mm. (standard) and Ra 0.005 mm., which is the standard glass we use in our production.

This Ra 0.005 mm. flatness, is specially selected glass, as is thickness Class #1.5H or 170 micron glass, +/- 5 micron.

Your WillCo Wells B.V. 'Customer Service-Team'

WillCo-dish®KIT 'Series KIT-3522'

Dear Colleague,

You may need your WillCo-dish® Glass Bottom dishes to have a coating, for better adherence of your cells. We do
not administer coatings on the glass of our WillCo-dish®es, because it is not allowed to use liquids in the high standard
clean room, where we manufacture our WillCo-dish®es. Besides, we only rarely received an inquiry, for coated glass.

Over the past twenty years, by far the most of our well respected WillCo-dish®USERS, took care of their own coatings,
the reason why we offer you some coating procedures, below. We welcome your suggestions, to further support your
Colleagues.

Coating procedures for glass coverslips
1.: COLLAGEN,
2.: POLYLYSINE and POLYORNITHINE,
3.: FIBRONECTIN,
4.: LAMININ.

1.: COLLAGEN

Collagen may be used to coat glass coverslips for the growth of epithelial, endothelial and muscle cells, neurons, PC12
and CHO cell lines.
Type I collagen is most often used for coating substrates for cell culture, because it is easily obtainable from rat tails. For
short term cultures, collagen can be simply applied to glass coverslips and allowed to dry.
1. Dilute collagen solution 1:10 - 1:50 with 30% ethanol and spread over surface of sterile glass coverslip.
2. Air dry in a tissue culture hood.
3. Cells can be seeded directly on the collagen surface.
4. Collagen coating prepared in this way tends to detach from the glass in long-term cultures.

Collagen IV is the major constituent of basement membrane and is therefore a more physiological coating for the culture
of many cell types.
For long-term cultures, collagen I and IV can be applied to glass coverslips by first coating the glass with polylysine or
polyornithine. This provides a more stable collagen coating.
1. Prepare polylysine or polyornithine (MW of 30,000 - 70,000) at 0.1-1 mg/ml in 0.15 M borate buffer
    (pH 8.3). Filter sterilize.
2. Add enough solution to pool over surface of sterile glass coverslip.
3. Incubate 2-24 hours at room temperature.
4. Aspirate solution and wash coverslips 3 times with water.
5. Pool collagen solution, 100 ug/ml in water over surface of coverslip.
6. Incubate 4 - 16 hours.
7. Rinse once with media and seed with cells.

Alternatively, for long-term cultures, double layered collagen coatings can provide a stable coating.
1. Spread a couple of drops of sterile collagen I solution on the sterile glass coverslip.
2. Immediately neutralize for 2 minutes with ammonium hydroxide vapors by placing the dish of coverslips
    in a covered dish containing filter paper wet with concentrated ammonium hydroxide. This will cause
    the collagen to gel.
3. Wash coverslips twice with sterile water.
4. Gently spread a couple of drops of collagen over the surface of the gelled collagen and air dry.
5. Use within a few hours for cell culture.

Gelatin can also be used for the culture of some cell types including glial cells.
1. Dissolve 100 mg gelatin in 100 ml water (triple glass distilled or RO).
2. Autoclave to sterilize.
3. While hot, thoroughly mix gelatin solution.
4. Add enough solution to pool over surface of sterile glass coverslip.
5. Chill for 2-24 hours at 4oC.
6. Remove gelatin by aspiration and add sterile water.
7. Dishes can be stored for up to one week at 4oC.
    Remove water immediately before use for cell culture

2.: POLYLYSINE AND POLYORNITHINE:
Nearly all types of cells adhere to these polymers of basic amino acids. They are particularly useful for the culture of CNS
neurons.
The L- or D-isomers can be used for cell attachment, however, the D-isomer may be preferred because it is not subject to
breakdown by proteases released by cells.
1. Prepare polylysine or polyornithine (MW of 30,000 - 70,000) at 0.1-1 mg/ml in 0.15 M borate buffer  (pH 8.3). Filter
    sterilize.
2. Add enough solution to pool over surface of sterile glass coverslip.
3. Incubate 2-24 hours at room temperature.
4. Aspirate solution and wash coverslips 3 times with media or PBS.
5. Immediately add cell suspension or growth media.

3.: FIBRONECTIN:
Fibronectin is an extracellular matrix constituent use for the culture of endothelial cells, fibroblasts, neurons and CHO cells.
1. Stock solution can be prepared by dissolving 1 mg/ml fibronectin in PBS. Filter sterilize and freeze in aliquots.
2. Diluted stock solution to 50-100 ug/ml in basal medium or PBS.
3. Add enough solution to pool over surface of sterile glass coverslip.
4. Incubate for 30-45 min at room temperature.
5. Aspirate to remove fibronectin and rinse coverslips with media or PBS.
6. Immediately add cell suspension or growth media. Do not allow coating to dry.

4.: LAMININ:
Laminin is an extracellular matrix constituent used for the culture of neurons, epithelial cells, leukocytes, myoblasts and
CHO cells.
1. Stock solution can be prepared by dissolving 1 mg/ml laminin in PBS. Filter sterilize and freeze in aliquots.
2. Diluted stock solution to 10-100 ug/ml in basal medium or PBS.
3. Add enough solution to pool over surface of sterile glass coverslip.
4. Incubate several hours at room temperature.
5. Aspirate to remove laminin and rinse coverslips with media or PBS.
6. Immediately add cell suspension or growth media. Do not allow coating to dry.
7. Coating the glass coverslip first with polylysine or polyornithine and then laminin may increase the
    concentration of laminin applied using this method.