Intravital 2-Photon Microscopy of Diverse Cell Types in the Murine Tibia

Hasenberg, Anja; Otto, Lucas; Gunzer, Matthias

doi:10.1007/978-1-0716-1060-2_15

Cited by 2 publications

(4 citation statements)

References 33 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The transferred cells recognize their cognate antigen, become activated and proliferate together with the endogenous FV-specific CTLs of the recipient (18). We then performed intravital two-photon microscopy (12,18,(26)(27)(28) at 14 days post FV infection to visualize individual moving CTL in the bone marrow of hosts. We chose this time point because we previously described that CTL exhaustion and impairment of CTL motility start at 14 days post FV infection (12).…”

Section: Resultsmentioning

confidence: 99%

“…Intravital two-photon microscopy was carried out as previously described (12,18,(26)(27)(28). Two-photon microscopy was done using a Leica TCS SP8 MP microscope (Leica Microsystems, Mannheim, Germany) with HCX IRAPO L25×/0.95-NA water-immersion objective, two external hybrid reflected-light detectors (HyD), and two external photomultiplier tubes (PMT).…”

Section: Intravital Two-photon Microscopy and Movie Analysismentioning

confidence: 99%

See 1 more Smart Citation

PD-1 knockout on cytotoxic primary murine CD8+ T cells improves their motility in retrovirus infected mice

Mittermüller,

Otto,

Kilian

et al. 2024

Front. Immunol.

Self Cite

View full text Add to dashboard Cite

Cytotoxic T lymphocyte (CTL) motility is an important feature of effective CTL responses and is impaired when CTLs become exhausted, e.g. during chronic retroviral infections. A prominent T cell exhaustion marker is programmed cell death protein 1 (PD-1) and antibodies against the interaction of PD-1 and PD-ligand 1 (PD-L1) are known to improve CTL functions. However, antibody blockade affects all PD-1/PD-L1-expressing cell types, thus, the observed effects cannot be attributed selectively to CTLs. To overcome this problem, we performed CRISPR/Cas9 based knockout of the PD-1 coding gene PDCD1 in naïve Friend Retrovirus (FV)-specific CTLs. We transferred 1,000 of these cells into mice where they proliferated upon FV-infection. Using intravital two-photon microscopy we visualized CTL motility in the bone marrow and evaluated cytotoxic molecule expression by flow cytometry. Knockout of PDCD1 improved the CTL motility at 14 days post infection and enhanced the expression of cytotoxicity markers. Our data show the potential of genetic tuning of naive antiviral CTLs and might be relevant for future designs of improved T cell-mediated therapies.

show abstract

Section: Resultsmentioning

confidence: 99%

Section: Intravital Two-photon Microscopy and Movie Analysismentioning

confidence: 99%

PD-1 knockout on cytotoxic primary murine CD8+ T cells improves their motility in retrovirus infected mice

Mittermüller,

Otto,

Kilian

et al. 2024

Front. Immunol.

Self Cite

View full text Add to dashboard Cite

show abstract

“…Consequently, TPLSM has become a powerful tool for studying biological function in live tissue thereby offering many advantages over conventional imaging techniques. In comparison with one‐photon fluorescent microscopy techniques, fluorescent dyes or proteins in TPLSM are excited not by a single photon but by two photons of lower energy, which provide the required excitation energy upon near simultaneous absorption that effectively combines the energies of both photons 60–63 . Thus, the required wavelength to excite a given fluorophore in TPLSM is red‐shifted to roughly two times its excitation wavelength in one‐photon excitation 64–66 .…”

Section: Intravital Two‐photon Microscopymentioning

confidence: 99%

“…In comparison with one-photon fluorescent microscopy techniques, fluorescent dyes or proteins in TPLSM are excited not by a single photon but by two photons of lower energy, which provide the required excitation energy upon near simultaneous absorption that effectively combines the energies of both photons. [60][61][62][63] Thus, the required wavelength to excite a given fluorophore in TPLSM is red-shifted to roughly two times its excitation wavelength in one-photon excitation. 6 4-66 Since it is using excitation with redshifted wavelengths, which are less subject to scattering by tissue structures, TPLSM can penetrate tissues much more deeply than one-photon microscopy, thereby reaching several hundreds of µm in native samples.…”

Section: Intr Avital T Wo -Photon MI Croscopymentioning

confidence: 99%

Imaging innate immunity*

Grüneboom

Aust

Cibir

et al. 2021

Immunological Reviews

Self Cite

View full text Add to dashboard Cite

The use of microscopes for the study of immune cells dates back to the 19th century with the German Pathologist Rudolf Virchow being one of the most prominent leaders in the field early on. It was him who coined the term "Zellularpathologie" (cellular pathology 1 ), while the famous concept associated with this theory, "omnis cellula e cellula" (all cells are derived from previous cells) was probably not originally from him. 2 Virchow was one of the earliest who observed by microscopy that inflammatory cells isolated from a punctured hydrocele were motile. Considering the inflammatory condition where they were harvested from and when looking at the images in his publication it might well be that he was looking at neutrophil granulocytes or macrophages at the time. 3 Hence, this could be the first description of the motility of these cells. More well-known are the famous studies by the Ukranian scientist Ilya Metchnikoff 20 years

show abstract

Intravital 2-Photon Microscopy of Diverse Cell Types in the Murine Tibia

Cited by 2 publications

References 33 publications

PD-1 knockout on cytotoxic primary murine CD8+ T cells improves their motility in retrovirus infected mice

PD-1 knockout on cytotoxic primary murine CD8+ T cells improves their motility in retrovirus infected mice

Imaging innate immunity*

Contact Info

Product

Resources

About