A variety of flask sizes filled with phenol-red medium

T25 flask dimensions

Start your scale-up with a T25 flask that has the surface and ancillary options you're looking for in a tissue culture flask. Choose a cell culture flask from a collection that includes straight-neck or angled-neck options and surface treatment alternatives that help ensure your healthy tissue cells expand and grow in the flask of your choice.

Some useful numbers such as surface area and volumes of dissociation solutions are given below for T25 flasks.

Please note: The number of cells on a flask will vary with cell type. For this table, 0.7 x 106 HeLa cells were seeded and the confluent flask contained 2.8 x 106 cells.

T25 flasks have a surface area of 25 cm2
Dissociation reagentVolume of reagentVolume of growth medium
TrypLE Express enzyme3.0 mL3–5 mL
TrypLE Select (1x) enzyme3.0 mL
TrypLE Select (10x) enzyme0.3 mL diluted to 3.0mL in DPBS/1 mM EDTA
Versene reagent3.0 mL
Trypsin3.0 mL

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T25 flasks: EasYFlask products

Ergonomically designed for maximal access to the growth surface, this low-profile flask optimizes incubator space and features angled walls for complete viewing under microscopes. Additional features include multiple cap options, and sterilization by gamma irradiation with non-pyrogenic certification.

EasYFlask product features

T25 flask surfaces by cell culture types

To help ensure flexible, reproducible, and reliable results across every stage of cell culture, T25 flasks come in a range of formats, sizes, and surfaces. Please see below for guidance on product selection for specific cell types and desired cell culture ranges.

Surface selection guide

Untreated surfaces for non-adherent cell suspension cultures

Made with high-quality, optically clear virgin polystyrene, the Thermo Scientific Nunc non-treated flasks, with high-level sterility assurance, are ideal for suspension culture of non-adherent cells.

A BioLite surface treated flask with blue cap

Thermo Scientific BioLite T25 surface treatments for general cell culture research

Thermo Scientific BioLite cell culture treated flasks with filter cap and seal cap options suit a multitude of experimental needs while providing cost efficiency for a wide selection of applications in your cell culture routine.

Caps come in both vented and plug seal varieties.

T25 flasks with poly-d-lysine coating or collagen I coatings

Poly-D-lysine- (PDL) and collagen I-coated flasks are ideal for adherent cultures of primary cells that are finicky and have difficulties attaching to the growth surface.

PDL is an animal-free synthetic surface for adherent cultures or primary cells that are finicky and hard to attach, while collagen I, also excellent for finicky cells, is of animal origin. Selection of one surface over another depends on tissue type. Please review previous references for your specific cell line or, if necessary, test both.

Untreated surfaces for non-adherent cell suspension cultures

Made with high-quality, optically clear virgin polystyrene, the Thermo Scientific Nunc non-treated flasks, with high-level sterility assurance, are ideal for suspension culture of non-adherent cells.

A BioLite surface treated flask with blue cap

Thermo Scientific BioLite T25 surface treatments for general cell culture research

Thermo Scientific BioLite cell culture treated flasks with filter cap and seal cap options suit a multitude of experimental needs while providing cost efficiency for a wide selection of applications in your cell culture routine.

Caps come in both vented and plug seal varieties.

T25 flasks with poly-d-lysine coating or collagen I coatings

Poly-D-lysine- (PDL) and collagen I-coated flasks are ideal for adherent cultures of primary cells that are finicky and have difficulties attaching to the growth surface.

PDL is an animal-free synthetic surface for adherent cultures or primary cells that are finicky and hard to attach, while collagen I, also excellent for finicky cells, is of animal origin. Selection of one surface over another depends on tissue type. Please review previous references for your specific cell line or, if necessary, test both.

     

Related flask and cell culture resources

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  • Alternative methods for advanced cell harvesting

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Surface selection: Achieving optimal cell growth and harvesting

Get an overview of technologies used to modify cultureware surfaces to help you select the right surface for your application.

  • Surface modifications to improve cell adhesion
  • Characterization of Nunc cell culture surfaces by adhesion properties
  • Alternative methods for advanced cell harvesting

Watch now

Establishing a dynamic micro-gravity spheroid 3D culture system

Learn about how to build specific 3D cell spheroid models and platforms to answer the pressing and relevant questions in cancer research.

  • Discerning the advantages and disadvantages of 2D versus 3D cell culturing approaches
  • Understanding the method of rotating micro-gravity bioreactors as a means to produce long-term spheroid cultures
  • Implementing spheroid cultures in cancer research

Watch now


Cell culture cafe

Watch more cell culture webinars

Register for our next on-demand webinar event and sign up to receive everything we have on cell culture.

Visit the café

Featured article

Cell adhesion and growth on coated or modified glass or plastic surfaces

Growth substrates affect the adhesion, growth, morphology and differentiation of various cell types. However, the response to a given surface is cell type specific. Selection of a growth surface for cell culture should be based on cell culture performance and on application. In this technical bulletin, the growth of various cell types on plastic, soda lime glass and borosilicate coverglass was examined with surfaces that were either unmodified, coated with polylysine, or stably surface modified with non-biological reagents or electrical discharge. 

Download bulletin

Are you on the front lines of cancer research?