bioimagesUnknownDaisuke NanbaThe Journal of Cell Biologyhttps://www.ebi.ac.uk/biostudies/studies/S-JCBD-201409024JCB

Shown is a rotating two-cell human epidermal keratinocyte colony with involucrin expression score of 0 (5 min). Images were obtained at 5 minutes intervals for 30 min. Involucrin expression was categorized in 3 groups depending on the number of involucrin-expressing cells: IVL score 0 (no cells), IVL score 1 (single cell), and IVL score 2 (two cells).

Shown is a IVL/ITGA6 staining of non-rotating two-cell human epidermal keratinocyte colony with involucrin expression score of 1. Colonies were immediately fixed after time-lapse imaging and stained with anti-alpha6 integrin (red) and anti-involucrin (green). DNA was also stained with Hoechst (blue). Involucrin expression was categorized in 3 groups depending on the number of involucrin-expressing cells: IVL score 0 (no cells), IVL score 1 (single cell), and IVL score 2 (two cells).

Shown is a time-lapse imaging series of a progressively growing colony of human keratinocytes after 6 days of cultivation. Images were obtained at 10 minutes intervals for 3 hours.

Shown is a non-rotating two-cell human epidermal keratinocyte colony with involucrin expression score of 1 (20 min). Images were obtained at 5 minutes intervals for 30 min. Involucrin expression was categorized in 3 groups depending on the number of involucrin-expressing cells: IVL score 0 (no cells), IVL score 1 (single cell), and IVL score 2 (two cells).

Shown is a rotating two-cell human epidermal keratinocyte colony (20 min). Images were obtained at 5 minutes intervals for 40 min.

Shown is a rotating two-cell human epidermal keratinocyte colony (40 min). Images were obtained at 5 minutes intervals for 40 min.

Shown is a progressively growing colony of human keratinocytes after 6 days of cultivation (110 min). Images were obtained at 10 minutes intervals for 3 hours.

Shown is a non-rotating two-cell human epidermal keratinocyte colony with involucrin expression score of 1 (25 min). Images were obtained at 5 minutes intervals for 30 min. Involucrin expression was categorized in 3 groups depending on the number of involucrin-expressing cells: IVL score 0 (no cells), IVL score 1 (single cell), and IVL score 2 (two cells).

Shown is a rotating two-cell human epidermal keratinocyte colony with involucrin expression score of 0 (10 min). Images were obtained at 5 minutes intervals for 30 min. Involucrin expression was categorized in 3 groups depending on the number of involucrin-expressing cells: IVL score 0 (no cells), IVL score 1 (single cell), and IVL score 2 (two cells).

Shown is Hoechst staining of a rotating two-cell human epidermal keratinocyte colony after 5 days cultivation. Clonal growth of two-cell colonies was traced and proliferative capacity was evaluated by counting cell number in the colony after 6 days of cultivation with Hoechst 33325.

Shown is a progressively growing colony of human keratinocytes after 6 days of cultivation (50 min). Images were obtained at 10 minutes intervals for 3 hours.

Shown is a rotating two-cell human epidermal keratinocyte colony (35 min). Images were obtained at 5 minutes intervals for 40 min.

Shown is a rotating two-cell human epidermal keratinocyte colony (15 min). Images were obtained at 5 minutes intervals for 40 min.

Shown is a progressively growing colony of human keratinocytes after 6 days of cultivation (70 min). Images were obtained at 10 minutes intervals for 3 hours.

Shown is alpha6 integrin expression in a rotating two-cell human epidermal keratinocyte colony. After a time-lapse imaging series of 60 minutes, the colony was immediately fixed and stained for anti-alpha6 integrin antibody and Hoechst.

Shown is a progressively growing colony of human keratinocytes (15 min). Images were obtained at a 5 min-interval for 1 hour.

Shown is a non-rotating two-cell human epidermal keratinocyte colony (15 min). Images were obtained at 5 minutes intervals for 30 min. Clonal growth of two-cell colonies was traced and proliferative capacity was evaluated by counting cell number in the colony after 6 days of cultivation with Hoechst 33325.

Shown is a progressively growing colony of human keratinocytes (25 min). Images were obtained at a 5 min-interval for 1 hour.

Shown is a non-rotating two-cell human epidermal keratinocyte colony with involucrin expression score of 1 (30 min). Images were obtained at 5 minutes intervals for 30 min. Involucrin expression was categorized in 3 groups depending on the number of involucrin-expressing cells: IVL score 0 (no cells), IVL score 1 (single cell), and IVL score 2 (two cells).

Shown is a progressively growing colony of human keratinocytes (35 min). Images were obtained at a 5 min-interval for 1 hour.

Shown is a rotating two-cell human epidermal keratinocyte colony (30 min). Images were obtained at 5 minutes intervals for 30 min. Clonal growth of two-cell colonies was traced and proliferative capacity was evaluated by counting cell number in the colony after 6 days of cultivation with Hoechst 33325.

Shown is a confocal imaging series of alpha6 (red) beta4 (green) integrin expression in a growing human keratinocyte colony.

Shown is a rotating two-cell human epidermal keratinocyte colony (5 min). Images were obtained at 5 minutes intervals for 40 min.

Shown is a progressively growing colony of human keratinocytes after 6 days of cultivation (120 min). Images were obtained at 10 minutes intervals for 3 hours.

Shown is a progressively growing colony of human keratinocytes (45 min). Images were obtained at a 5 min-interval for 1 hour.

Shown is a rotating two-cell human epidermal keratinocyte colony (0 min). Images were obtained at 5 minutes intervals for 40 min.

Shown is a non-rotating two-cell human epidermal keratinocyte colony (25 min). Images were obtained at 5 minutes intervals for 30 min. Clonal growth of two-cell colonies was traced and proliferative capacity was evaluated by counting cell number in the colony after 6 days of cultivation with Hoechst 33325.<br />

Shown is a rotating two-cell human epidermal keratinocyte colony with involucrin expression score of 0 (25 min). Images were obtained at 5 minutes intervals for 30 min. Involucrin expression was categorized in 3 groups depending on the number of involucrin-expressing cells: IVL score 0 (no cells), IVL score 1 (single cell), and IVL score 2 (two cells).

Shown is a progressively growing colony of human keratinocytes after 6 days of cultivation (40 min). Images were obtained at 10 minutes intervals for 3 hours.

Shown is the expression of alpha6 (red) and beta4 integrin (green) in a progressively growing human keratinocyte colony. Cells were treated with 0.5% Triton X-100 after fixation.

Shown is a non-rotating two-cell human epidermal keratinocyte colony with involucrin expression score of 1 (0 min). Images were obtained at 5 minutes intervals for 30 min. Involucrin expression was categorized in 3 groups depending on the number of involucrin-expressing cells: IVL score 0 (no cells), IVL score 1 (single cell), and IVL score 2 (two cells).

Shown is a progressively growing colony of human keratinocytes (55 min). Images were obtained at a 5 min-interval for 1 hour.

Shown is a non-rotating two-cell human epidermal keratinocyte colony with involucrin expression score of 1 (5 min). Images were obtained at 5 minutes intervals for 30 min. Involucrin expression was categorized in 3 groups depending on the number of involucrin-expressing cells: IVL score 0 (no cells), IVL score 1 (single cell), and IVL score 2 (two cells).

Shown is a progressively growing colony of human keratinocytes after 6 days of cultivation (80 min). Images were obtained at 10 minutes intervals for 3 hours.

Shown is a progressively growing colony of human keratinocytes after 6 days of cultivation (150 min). Images were obtained at 10 minutes intervals for 3 hours.

Shown is a progressively growing colony of human keratinocytes after 6 days of cultivation (10 min). Images were obtained at 10 minutes intervals for 3 hours.

Shown is Hoechst staining of a rotating two-cell human epidermal keratinocyte colony. After a time-lapse imaging series of 60 minutes, the colony was immediately fixed and stained for anti-alpha6 integrin antibody and Hoechst.

Shown is a time-lapse imaging series of a growing human keratinocyte colony (average speed = 34.2). Images were obtained at 5 minutes intervals for 3 hours.

Shown is a rotating two-cell human epidermal keratinocyte colony (25 min). Images were obtained at 5 minutes intervals for 40 min.

Shown is a rotating two-cell human epidermal keratinocyte colony with involucrin expression score of 0 (0 min). Images were obtained at 5 minutes intervals for 30 min. Involucrin expression was categorized in 3 groups depending on the number of involucrin-expressing cells: IVL score 0 (no cells), IVL score 1 (single cell), and IVL score 2 (two cells).

Shown is the colony forming-efficiency of an isolated growing colony of human keratinocytes (% of terminal colonies = 0). After time-lapse imaging, the colony was isolated, trypsinized, and seeded into a 10 cm dish to evaluate growth potential.

Shown is a time-lapse imaging series of a growing human keratinocyte colony (average speed = 37.8). Images were obtained at 5 minutes intervals for 3 hours.

Shown is a rotating two-cell human epidermal keratinocyte colony with involucrin expression score of 0 (30 min). Images were obtained at 5 minutes intervals for 30 min. Involucrin expression was categorized in 3 groups depending on the number of involucrin-expressing cells: IVL score 0 (no cells), IVL score 1 (single cell), and IVL score 2 (two cells).

Shown is Hoechst staining of a non-rotating two-cell human epidermal keratinocyte colony after 5 days cultivation. Clonal growth of two-cell colonies was traced and proliferative capacity was evaluated by counting cell number in the colony after 6 days of cultivation with Hoechst 33325.

Shown is a non-rotating two-cell human epidermal keratinocyte colony with involucrin expression score of 2 (0 min). Images were obtained at 5 minutes intervals for 30 min. Involucrin expression was categorized in 3 groups depending on the number of involucrin-expressing cells: IVL score 0 (no cells), IVL score 1 (single cell), and IVL score 2 (two cells).

Shown is a progressively growing colony of human keratinocytes after 6 days of cultivation (170 min). Images were obtained at 10 minutes intervals for 3 hours.

Shown is the colony forming-efficiency of an isolated growing colony of human keratinocytes % of terminal colonies = 52.1). After time-lapse imaging, the colony was isolated, trypsinized, and seeded into a 10 cm dish to evaluate growth potential.

Shown is a progressively growing colony of human keratinocytes (30 min). Images were obtained at a 5 min-interval for 1 hour.

Shown is a progressively growing colony of human keratinocytes (20 min). Images were obtained at a 5 min-interval for 1 hour.

Shown is a rotating two-cell human epidermal keratinocyte colony (15 min). Images were obtained at 5 minutes intervals for 30 min. Clonal growth of two-cell colonies was traced and proliferative capacity was evaluated by counting cell number in the colony after 6 days of cultivation with Hoechst 33325.

Shown is a progressively growing colony of human keratinocytes after 6 days of cultivation (0 min). Images were obtained at 10 minutes intervals for 3 hours.

Shown is phase contrast image of a non-rotating two-cell human epidermal keratinocyte colony after 5 days cultivation. Clonal growth of two-cell colonies was traced and proliferative capacity was evaluated by counting cell number in the colony after 6 days of cultivation with Hoechst 33325.

Shown is a progressively growing colony of human keratinocytes after 6 days of cultivation (160 min). Images were obtained at 10 minutes intervals for 3 hours.

Shown is a progressively growing colony of human keratinocytes (0 min). Images were obtained at a 5 min-interval for 1 hour.

Shown is a rotating two-cell human epidermal keratinocyte colony (20 min). Images were obtained at 5 minutes intervals for 30 min. Clonal growth of two-cell colonies was traced and proliferative capacity was evaluated by counting cell number in the colony after 6 days of cultivation with Hoechst 33325.

Shown is an electron micrograph of a growing colony of human epidermal keratinocytes (x20000).

Shown is an electron micrograph of a growing colony of human epidermal keratinocytes (x50000).

Shown is a rotating two-cell human epidermal keratinocyte colony with involucrin expression score of 0 (20 min). Images were obtained at 5 minutes intervals for 30 min. Involucrin expression was categorized in 3 groups depending on the number of involucrin-expressing cells: IVL score 0 (no cells), IVL score 1 (single cell), and IVL score 2 (two cells).

Shown is a progressively growing colony of human keratinocytes (10 min). Images were obtained at a 5 min-interval for 1 hour.

Shown is a rotating two-cell human epidermal keratinocyte colony (5 min). Images were obtained at 5 minutes intervals for 30 min. Clonal growth of two-cell colonies was traced and proliferative capacity was evaluated by counting cell number in the colony after 6 days of cultivation with Hoechst 33325.

Shown is a non-rotating two-cell human epidermal keratinocyte colony with involucrin expression score of 2 (25 min). Images were obtained at 5 minutes intervals for 30 min. Involucrin expression was categorized in 3 groups depending on the number of involucrin-expressing cells: IVL score 0 (no cells), IVL score 1 (single cell), and IVL score 2 (two cells).

Shown is a rotating two-cell human epidermal keratinocyte colony (10 min). Images were obtained at 5 minutes intervals for 40 min.

Shown is a rotating two-cell human epidermal keratinocyte colony (30 min). Images were obtained at 5 minutes intervals for 40 min.

Shown is a rotating two-cell human epidermal keratinocyte colony (0 min). Images were obtained at 5 minutes intervals for 30 min. Clonal growth of two-cell colonies was traced and proliferative capacity was evaluated by counting cell number in the colony after 6 days of cultivation with Hoechst 33325.

Shown is a time-lapse imaging series of a growing human keratinocyte colony (average speed = 27.7). Images were obtained at 5 minutes intervals for 3 hours.

Shown is a progressively growing colony of human keratinocytes (50 min). Images were obtained at a 5 min-interval for 1 hour.

Shown is a IVL/ITGA6 staining of non-rotating two-cell human epidermal keratinocyte colony with involucrin expression score of 2. Colonies were immediately fixed after time-lapse imaging and stained with anti-alpha6 integrin (red) and anti-involucrin (green). DNA was also stained with Hoechst (blue). Involucrin expression was categorized in 3 groups depending on the number of involucrin-expressing cells: IVL score 0 (no cells), IVL score 1 (single cell), and IVL score 2 (two cells).

Shown is a non-rotating two-cell human epidermal keratinocyte colony with involucrin expression score of 2 (30 min). Images were obtained at 5 minutes intervals for 30 min. Involucrin expression was categorized in 3 groups depending on the number of involucrin-expressing cells: IVL score 0 (no cells), IVL score 1 (single cell), and IVL score 2 (two cells).

Shown is a rotating two-cell human epidermal keratinocyte colony with involucrin expression score of 0 (15 min). Images were obtained at 5 minutes intervals for 30 min. Involucrin expression was categorized in 3 groups depending on the number of involucrin-expressing cells: IVL score 0 (no cells), IVL score 1 (single cell), and IVL score 2 (two cells).

Shown is a rotating two-cell human epidermal keratinocyte colony (25 min). Images were obtained at 5 minutes intervals for 30 min. Clonal growth of two-cell colonies was traced and proliferative capacity was evaluated by counting cell number in the colony after 6 days of cultivation with Hoechst 33325.

Shown is a progressively growing colony of human keratinocytes after 6 days of cultivation (140 min). Images were obtained at 10 minutes intervals for 3 hours.

Shown is a progressively growing colony of human keratinocytes (40 min). Images were obtained at a 5 min-interval for 1 hour.

Shown is a rotating two-cell human epidermal keratinocyte colony (10 min). Images were obtained at 5 minutes intervals for 30 min. Clonal growth of two-cell colonies was traced and proliferative capacity was evaluated by counting cell number in the colony after 6 days of cultivation with Hoechst 33325.

Shown is a non-rotating two-cell human epidermal keratinocyte colony (30 min). Images were obtained at 5 minutes intervals for 30 min. Clonal growth of two-cell colonies was traced and proliferative capacity was evaluated by counting cell number in the colony after 6 days of cultivation with Hoechst 33325.

Shown is a non-rotating two-cell human epidermal keratinocyte colony with involucrin expression score of 2 (5 min). Images were obtained at 5 minutes intervals for 30 min. Involucrin expression was categorized in 3 groups depending on the number of involucrin-expressing cells: IVL score 0 (no cells), IVL score 1 (single cell), and IVL score 2 (two cells).

Shown is a progressively growing colony of human keratinocytes after 6 days of cultivation (20 min). Images were obtained at 10 minutes intervals for 3 hours.

Shown is a non-rotating two-cell human epidermal keratinocyte colony (0 min). Images were obtained at 5 minutes intervals for 30 min. Clonal growth of two-cell colonies was traced and proliferative capacity was evaluated by counting cell number in the colony after 6 days of cultivation with Hoechst 33325.

Shown is a progressively growing colony of human keratinocytes after 6 days of cultivation (130 min). Images were obtained at 10 minutes intervals for 3 hours.

Shown is a progressively growing colony of human keratinocytes (5 min). Images were obtained at a 5 min-interval for 1 hour.

Shown is a progressively growing colony of human keratinocytes after 6 days of cultivation (30 min). Images were obtained at 10 minutes intervals for 3 hours.

Shown is the expression of alpha6 and beta4 integrin in a growing human keratinocyte colony. A maximum intensity projection image of immunofluorescence against alpha6 (red) and beta4 (green) integrin in a growing colony by confocal microscopy.

Shown is the colony forming-efficiency of an isolated growing colony of human keratinocytes (% of terminal colonies = 5.61). After time-lapse imaging, the colony was isolated, trypsinized, and seeded into a 10 cm dish to evaluate growth potential.

Shown is the alpha6 integrin expression in a growing human keratinocyte colony. After time-lapse imaging, the colony was immediately fixed and stained with anti-alpha6 integrin antibody (green) and Hoechst (blue).

Shown is a non-rotating two-cell human epidermal keratinocyte colony with involucrin expression score of 2 (20 min). Images were obtained at 5 minutes intervals for 30 min. Involucrin expression was categorized in 3 groups depending on the number of involucrin-expressing cells: IVL score 0 (no cells), IVL score 1 (single cell), and IVL score 2 (two cells).

Shown is phase contrast image of a rotating two-cell human epidermal keratinocyte colony after 5 days cultivation. Clonal growth of two-cell colonies was traced and proliferative capacity was evaluated by counting cell number in the colony after 6 days of cultivation with Hoechst 33325.

Shown is a progressively growing colony of human keratinocytes after 6 days of cultivation (90 min). Images were obtained at 10 minutes intervals for 3 hours.

Shown is a non-rotating two-cell human epidermal keratinocyte colony (20 min). Images were obtained at 5 minutes intervals for 30 min. Clonal growth of two-cell colonies was traced and proliferative capacity was evaluated by counting cell number in the colony after 6 days of cultivation with Hoechst 33325.

Shown is a non-rotating two-cell human epidermal keratinocyte colony (10 min). Images were obtained at 5 minutes intervals for 30 min. Clonal growth of two-cell colonies was traced and proliferative capacity was evaluated by counting cell number in the colony after 6 days of cultivation with Hoechst 33325.

Shown is a IVL/ITGA6 staining of a rotating two-cell human epidermal keratinocyte colony with involucrin expression score of 0. Colonies were immediately fixed after time-lapse imaging and stained with anti-alpha6 integrin (red) and anti-involucrin (green). DNA was also stained with Hoechst (blue). Involucrin expression was categorized in 3 groups depending on the number of involucrin-expressing cells: IVL score 0 (no cells), IVL score 1 (single cell), and IVL score 2 (two cells).

Shown is a progressively growing colony of human keratinocytes after 6 days of cultivation (100 min). Images were obtained at 10 minutes intervals for 3 hours.

Shown is a non-rotating two-cell human epidermal keratinocyte colony with involucrin expression score of 2 (10 min). Images were obtained at 5 minutes intervals for 30 min. Involucrin expression was categorized in 3 groups depending on the number of involucrin-expressing cells: IVL score 0 (no cells), IVL score 1 (single cell), and IVL score 2 (two cells).

Shown is a non-rotating two-cell human epidermal keratinocyte colony with involucrin expression score of 2 (15 min). Images were obtained at 5 minutes intervals for 30 min. Involucrin expression was categorized in 3 groups depending on the number of involucrin-expressing cells: IVL score 0 (no cells), IVL score 1 (single cell), and IVL score 2 (two cells).

Shown is a non-rotating two-cell human epidermal keratinocyte colony (5 min). Images were obtained at 5 minutes intervals for 30 min. Clonal growth of two-cell colonies was traced and proliferative capacity was evaluated by counting cell number in the colony after 6 days of cultivation with Hoechst 33325.

Shown is a progressively growing colony of human keratinocytes after 6 days of cultivation (60 min). Images were obtained at 10 minutes intervals for 3 hours.

Shown is a Hoechst staining of a growing human keratinocyte colony. After time-lapse imaging, the colony was immediately fixed and stained with anti-alpha6 integrin antibody (green) and hoechst (blue).

Shown is a non-rotating two-cell human epidermal keratinocyte colony with involucrin expression score of 1 (10 min). Images were obtained at 5 minutes intervals for 30 min. Involucrin expression was categorized in 3 groups depending on the number of involucrin-expressing cells: IVL score 0 (no cells), IVL score 1 (single cell), and IVL score 2 (two cells).

Shown is a non-rotating two-cell human epidermal keratinocyte colony with involucrin expression score of 1 (15 min). Images were obtained at 5 minutes intervals for 30 min. Involucrin expression was categorized in 3 groups depending on the number of involucrin-expressing cells: IVL score 0 (no cells), IVL score 1 (single cell), and IVL score 2 (two cells).

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