time lapse videos  (Oxford Instruments)

Journal: bioRxiv

Article Title: Dual lineages of Langerhans cells cooperate to restore the immune barrier after skin injury

doi: 10.64898/2026.05.25.727646

Figure Lengend Snippet: a , Revisit multi-photon in-vivo microscopy images of a 1 mm wound from the same mouse. Image shows x-y view of epithelial cells (red nuclei) and LCs (green) in the epidermis. Dashed line indicates initial wound boundary. Left : day of wound induction (day 0). Middle : 5 days after wound induction. Right : zoomed view of the wound center at day 5. b , Time-lapse image of epithelial cells (red nuclei) and LCs (green) 2 days after wound induction. Dashed line, initial wound boundary. Solid line, basal membrane separating epidermis from dermis. Top : x-y view. Bottom : x-z view shows the epidermis (red) and dermis (collagen SHG, blue). c , Imaris track analysis of LCs ( b ) 2 days after wound induction. Colors project time (blue, 0h; red, 6h). Top : x-y view. Bottom : x-z view. d , Top : zoomed migration tracks from ( c ). The green frame is from the wound leading-edge epithelial migration zone, and the teal frame is from the epithelial proliferation zone 3 . Middle : time-lapse frames show the movement of individually colored LCs across 6 hours. Epithelial cells nuclei in gray. Other LCs in white. Bottom : vector arrows show the general movement direction of the matching color LC. e , Mean total displacement of individual LC tracks over 6h plotted as a function of distance from the wound. f , Mean track displacement in the x axis of individual LC tracks over 6h plotted as a function of distance from the wound. Calculated by comparing the start and end values in the x axis of each track. Positive change indicates movement towards the wound. g , Imaris cell count analysis of LCs (spots) at day 0 ( left ) and 5 days after wound induction ( right ). Dashed lines separate LCs into 3 zones: wound (yellow spots), near (0-400 µm from the wound edge, green spots), and far (400-700 µm from the wound edge, teal spots). Epithelial cells’ nuclei are shown in gray. h , Mean LC number comparing cell density between day 0 and 5 days after wound induction according to the 3 zones established in ( g ). i , Mean LC number comparing the cell density change from the addition of the wound and near zones between day 0 and 5 days after wound induction ( left bars ). Total change in LC density across all 3 zones between day 0 and 5 days after wound induction ( right bars ). h-i , Data analyzed using paired two-way ANOVA; n =3 mice; data are mean ±s.d. with each dot representing individual mice. *** P < 0.001, **** P < 0.0001. e-f , Imaging performed 2 days after wound induction. The displacements of migrating cell tracks were averaged every 100 µm from the initial wound. n =3 mice; data are mean ±s.d. Representative images from 3 mice. Scale bars, 100 µm.

Article Snippet: For time-lapse videos, Imaris was used to track cells and obtain xyz coordinates from individual cells over time.

Techniques: In Vivo, Microscopy, Membrane, Migration, Plasmid Preparation, Cell Characterization, Imaging

Journal: bioRxiv

Article Title: Dual lineages of Langerhans cells cooperate to restore the immune barrier after skin injury

doi: 10.64898/2026.05.25.727646

Figure Lengend Snippet: a , Time-lapse x-y view of epithelial cells (red nuclei) and LCs (green) 2 days after wound induction. Top : control mouse. Bottom : KTC- Rac1 KO mouse. b , Imaris x-y view track analysis of LCs ( a ) 2 days after wound induction. Colors project time (blue, 0h; red, 6h). Top : control mouse. Bottom : KTC- Rac1 KO mouse. Right : zoomed migration tracks from near the wound edge. a, b Dashed line indicates initial wound boundary. Representative images from 3 mice. Scale bars, 100 µm. c , Mean total displacement of individual epithelial cells tracks from control and KTC- Rac1 KO mice over 6h plotted as a function of distance from the wound. d , Mean total displacement of individual LC tracks from control and KTC- Rac1 KO mice over 6h plotted as a function of distance from the wound. c-d , Imaging performed 2 days after wound induction. The displacements of migrating cell tracks were averaged every 100 µm from the initial wound. Data analyzed using unpaired two-way ANOVA; n =3 mice; data are mean ±s.d. * P < 0.05. e , In-vivo microscopy images shows x-y view of epithelial cells (red nuclei) and LCs (green) in the epidermis 5 days after wound induction. Dashed line indicates initial wound boundary. Left : control mouse and LC- Rac1 KO mouse. Right : zoomed view of the wound center from 5 days after wound induction. Representative images from 4 mice. Scale bars, 200 µm. f , Mean LC number inside the wound epidermis from control and LC- Rac1 KO mice. Imaging was performed 5 days after wound induction. LC density normalized to the individual mouse wound area quantified. Data analyzed using unpaired two-tailed t -test; n =4 mice; data are mean ±s.d. with each dot representing individual mice. **** P < 0.0001.

Article Snippet: For time-lapse videos, Imaris was used to track cells and obtain xyz coordinates from individual cells over time.

Techniques: Control, Migration, Imaging, In Vivo, Microscopy, Two Tailed Test

Journal: bioRxiv

Article Title: Dual lineages of Langerhans cells cooperate to restore the immune barrier after skin injury

doi: 10.64898/2026.05.25.727646

Figure Lengend Snippet: a , Revisit multi-photon in-vivo microscopy images of a 1 mm wound from the same mouse. Images show x-y view of epithelial cells (red nuclei) and LCs (green) in the epidermis at 0, 5, 15 days, and 6 weeks after wound induction. Dashed line indicates initial wound boundary. Left : zoomed view of the wound center. Right : revisit images showing the classification of several zones according to their distance from the wound: wound, near, middle, and far. Representative images from 6 mice. Scale bars, 100 µm. b , Mean LC number comparing cell density changes within designated zones established in ( a ) at 0, 5, 15 days, and 6 weeks after wound induction. c , Timeline of LC number comparing cell density changes between the wound and near zones. b-c , Data analyzed using unpaired two-way ANOVA; n =6 mice; data are mean ±s.d. * P < 0.05, ** P < 0.01. d , Time-lapse image in x-y view of proliferative LCs (green) at 7 days after wound induction. Dermis SHG collagen is shown in gray. Red dashed circles indicate diving LCs. e , Time-lapse frames from yellow highlighted area in ( d ) show LC division across 5 hours. White arrows indicate actively diving LC. d-e , Representative images from at least 2 mice. White dashed circle/line indicates initial wound boundary. Scale bars, 50 µm. f , Confocal immunofluorescent images of cell proliferation at the wound epidermis 7 days after wound induction. Images show x-y view of LCs (green, MHC-II), proliferation (red, Ki67), and cell nuclei (blue, DAPI). Left : composite image. Middle : MHC-II and Ki67 positive cells. Right : zoomed example of proliferative LC. MHC-II, major histocompatibility complex class II. White arrows show proliferative cells. White arrowheads show proliferative LCs. g , Timeline of mean proliferative (Ki67+) cell density at the wound during healing and in homeostasis. h , Timeline of percentage of proliferative LCs (MHC-II+Ki67+) at the wound during healing and in homeostasis. i , Confocal immunofluorescent images of cell apoptosis at the wound epidermis 3 weeks after wound induction. Images show x-y view of LCs (green, MHC-II), apoptosis (red, CC3), and cell nuclei (blue, DAPI). Left : composite image. Middle : MHC-II and CC3 positive cells. Right : zoomed example of apoptotic LC. CC3, cleaved caspase-3. The white arrow shows an apoptotic cell. White arrowhead shows an apoptotic LC. f , i Representative images from 4 mice. The white dashed line indicates the initial wound boundary. Scale bars, 50 µm. j , Mean number of apoptotic cells (CC3+) and apoptotic LCs (CC3+MHC-II+) at the wound 3 weeks after wound induction. k , Percentage of apoptotic LCs (CC3+MHC-II+) during homeostasis and at 3 weeks after wound induction. g - h , j - k , Wound area quantified 0.49 mm 2 per mouse. Data analyzed using unpaired one-way ANOVA; n ≥3 mice; data are mean ±s.d. with each dot representing individual mice. * P < 0.05, **** P < 0.0001. l , Time-lapse frames show in vivo LC (green) apoptosis across 4 hours. The white dash circle highlights apoptotic LC. n =1 mouse. Scale bar, 50 µm.

Article Snippet: For time-lapse videos, Imaris was used to track cells and obtain xyz coordinates from individual cells over time.

Techniques: In Vivo, Microscopy, Immunopeptidomics

Journal: bioRxiv

Article Title: Dual lineages of Langerhans cells cooperate to restore the immune barrier after skin injury

doi: 10.64898/2026.05.25.727646

Figure Lengend Snippet: a , Heatmap of normalized log2 fold change of chemokine receptor gene expression from homeostatic epithelial (KTC) and Langerhans cells (LC). Cells isolated through FACS and sequenced through bulk RNA-seq. Each column represents an independent sample, and each row is assigned to a specific gene. Red indicates maximum expression and blue indicates minimum expression. b , Experimental design for drug treatment. Starting on wound induction day, drug was injected once a day intradermally at the ear near the wound site. Control mice received vehicle (1% DMSO) injections. Wounds were imaged at wound closure (5 days after wound induction), and candidate drugs were further analyzed for migration dynamics through time-lapse at 2 days after wound induction. c , Mean LC number comparing cell density at the wound in response to drug treatment. Imaging was performed 5 days after wound induction. LC density normalized to the individual mouse wound area was quantified. Data analyzed using unpaired one-way ANOVA; n ≥4 mice; data are mean ±s.d. with each dot representing individual mice. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001. d , In-vivo microscopy images show x-y view of epithelial cells (red nuclei) and LCs (green) in the epidermis 5 days after wound induction. Top : control mouse (1% DMSO). Middle : CXCR2-inhibited mouse (Danirixin). Bottom : CXCR2-inhibited mouse (SB225002). Right : zoomed view of the wound center matching the image on the left. Dashed line indicates initial wound boundary. Representative images from 5 mice. Scale bars, 100 µm. e , Confocal immunofluorescent images of CXCR2 expression at the epidermis during homeostasis and 2 days after wound induction. Images show x-y view of LCs (green, MHC-II), CXCR2 (red), and cell nuclei (blue, DAPI). Right : zoomed view in composite, green channel only, and red channel only. Dashed line indicates initial wound boundary. Representative images from 3 mice. Scale bars, 25 µm. f , qRT-PCR gene expression analysis of CXCR2 ligands in the skin during homeostasis (control) and 2 days after wound induction. Data analyzed using multiple unpaired two-tailed t -test; n =5 mice; data are mean ±s.d. with each dot representing individual mice. ** P < 0.01.

Article Snippet: For time-lapse videos, Imaris was used to track cells and obtain xyz coordinates from individual cells over time.

Techniques: Gene Expression, Isolation, RNA Sequencing, Expressing, Injection, Control, Migration, Imaging, In Vivo, Microscopy, Quantitative RT-PCR, Two Tailed Test

Journal: bioRxiv

Article Title: Dual lineages of Langerhans cells cooperate to restore the immune barrier after skin injury

doi: 10.64898/2026.05.25.727646

Figure Lengend Snippet: a , Time-lapse x-y view of epithelial cells (red nuclei) and LCs (green) 2 days after wound induction. Top : control mouse 1% DMSO. Bottom : drug-treated mouse CXCR2 (Danirixin) as shown in . Dashed line indicates initial wound boundary. Representative images from 3 mice. Scale bars, 100 µm. b , Imaris x-y view track analysis of LCs ( a ) 2 days after wound induction. Colors project time (blue, 0h; red, 6h). Top : control mouse 1% DMSO. Bottom : drug-treated mouse CXCR2 (Danirixin). Right : zoomed migration tracks from near the wound edge. c , Mean total displacement of individual epithelial cells tracks from control and CXCR2-inhibited mice over 6h plotted as a function of distance from the wound. d , Mean total displacement of individual LC tracks from control and CXCR2-inhibited mice over 6h plotted as a function of distance from the wound. e , Mean track displacement in the x axis of individual LC tracks from control and CXCR2-inhibited mice over 6h plotted as a function of distance from the wound. Calculated by comparing the start and end values on the x axis of each track. Positive change indicates movement towards the wound. c-e , Imaging performed 2 days after wound induction. The displacements of migrating cell tracks were averaged every 100 µm from the initial wound. Data analyzed using unpaired two-way ANOVA; n =3 mice; data are mean ±s.d. * P < 0.05, **** P < 0.0001. f , Revisit multi-photon in-vivo microscopy images of a 1 mm wound from the same mouse. Images show x-y view of epithelial cells (dim red nuclei), embryonic LCs (orange/yellow), and progenitor-derived LCs (green) in the epidermis at 5 ( Left ) and 17 ( Middle ) days after wound induction. Day 17 LCs at the wound quantified using Imaris spots analysis ( Right ). Top : control mouse 1% DMSO. Bottom : drug-treated mouse CXCR2 (Danirixin). Dashed line indicates initial wound boundary. Representative images from at least 3 mice. Scale bar, 100 µm. g , Mean LC number comparing cell density changes among embryonic LCs (eLC) and monocyte-derived LCs (mLC) in response to CXCR2 inhibition. h , Percentage ratio between eLCs and mLCs at the wound epidermis in response to CXCR2 inhibition. g-h , Imaging performed 17 days after wound induction. LC density normalized to the individual mouse wound area was quantified. Data analyzed using unpaired two-way ANOVA; n ≥3 mice; data are mean ±s.d. * P < 0.05.

Article Snippet: For time-lapse videos, Imaris was used to track cells and obtain xyz coordinates from individual cells over time.

Techniques: Control, Migration, Imaging, In Vivo, Microscopy, Derivative Assay, Inhibition