MPScope: A versatile software suite for multiphoton microscopy

Nguyen, Quoc-Thang; Tsai, Philbert S.; Kleinfeld, David

doi:10.1016/j.jneumeth.2006.03.001

Cited by 99 publications

(81 citation statements)

References 17 publications

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“…The galvanometric mirrors, stage, and PMT are controlled and synchronized by MPScan software. 24 Both the lateral and axial point spread functions (respective FWHM of 0.87 AE 0.05 μm and 3.4 AE 0.4 μm for THG; 0.59 AE 0.08 μm and 2.6 AE 0.4 μm for TPAF) remain constant in the range of depths used in our experiments, indicating little to no degradation from spherical aberration.…”

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confidence: 53%

Determination of scattering properties and damage thresholds in tissue using ultrafast laser ablation

Martin

Ben‐Yakar

2016

J. Biomed. Opt

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confidence: 53%

Determination of scattering properties and damage thresholds in tissue using ultrafast laser ablation

Martin

Ben‐Yakar

2016

J. Biomed. Opt

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“…Animals were imaged (32) using a custom twophoton microscope (33) that employed photon counting detection (34) and was controlled by MPScan software (35). We further used a 10× 0.…”

Section: Methodsmentioning

confidence: 99%

Fluctuating and sensory-induced vasodynamics in rodent cortex extend arteriole capacity

Drew

Shih

Kleinfeld

2011

Proc. Natl. Acad. Sci. U.S.A.

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261

388

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Neural activity in the brain is followed by localized changes in blood flow and volume. We address the relative change in volume for arteriole vs. venous blood within primary vibrissa cortex of awake, head-fixed mice. Two-photon laser-scanning microscopy was used to measure spontaneous and sensory evoked changes in flow and volume at the level of single vessels. We find that arterioles exhibit slow (<1 Hz) spontaneous increases in their diameter, as well as pronounced dilation in response to both punctate and prolonged stimulation of the contralateral vibrissae. In contrast, venules dilate only in response to prolonged stimulation. We conclude that stimulation that occurs on the time scale of natural stimuli leads to a net increase in the reservoir of arteriole blood. Thus, a "bagpipe" model that highlights arteriole dilation should augment the current "balloon" model of venous distension in the interpretation of fMRI images.ocalized changes in the flow and volume of oxygenated blood in the brain are commonly used as a correlate of heightened neural activity. For two important imaging modalities, blood oxygen level-dependent functional magnetic resonance imaging (BOLD fMRI) (1) and intrinsic optical signal imaging (IOS) (2), the signals are generated by a complex interplay of the rate of oxidative metabolism, the flux of blood in the underlying angioarchitecture, and changes in vascular volume (3). The locus for the increase in vascular volume that follows sensory stimulation (i.e., arterioles or venules) is an enduring controversy that bears directly on interpreting and quantifying fMRI signals (4-6). To resolve this question, we used in vivo two-photon laserscanning microscopy to image spontaneous and sensory evoked vascular dynamics in the vibrissa area of parietal cortex of awake, head-fixed mice. All data were collected through a reinforced thin-skull window (7) (Fig. 1A); this method obviates potential complications from inflammation or changes in cranial pressure that may occur with a craniotomy (8). ResultsImages of the pial surface and measurements of the diameter of the lumen of surface arterioles and venules were performed while the mouse sat passively (Fig. 1B). Dilations greater than 5% of the baseline diameter occurred with frequencies of 0.07 ± 0.05 Hz (mean ± SD, n = 118 arteries in six mice). The diameters of short segments of arterioles (red in Fig. 1B) exhibited relatively large spontaneous increases in the spectral range between 0.1 Hz and 1 Hz ( Fig. 1 C and D). The peak amplitude of these fluctuations was 23% ± 10% of the initial vessel diameter across all arterioles, with instances of a 50% increases in diameter. Further, these lowfrequency oscillations were strongly coherent and synchronous over a distance of several hundred micrometers across cortex (198 pairs of vessels, in six mice, that were separated by 20-315 μm; slope of coherence = 0.001 μm −1 [nonsignificant (NS)] and slope of phase shift = 0.0009 rad/μm

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“…Images were collected using a 2-photon laser scanning microscope of custom design (49) that was controlled by MPScope software (50). The blood serum was labeled using 0.1 ml of 2 MDa fluorescein dextran (FD2000S; Sigma-Aldrich) prepared at a concentration of 2.5% (w/v) in physiological saline and delivered through a tail-vein injection with 0.1 ml supplements as required.…”

Section: Methodsmentioning

confidence: 99%

The glial cell response is an essential component of hypoxia-induced erythropoiesis in mice

Weidemann¹,

Kerdiles²,

Knaup³

et al. 2009

J. Clin. Invest.

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A key adaptation to environmental hypoxia is an increase in erythropoiesis, driven by the hormone erythropoietin (EPO) through what is traditionally thought to be primarily a renal response. However, both neurons and astrocytes (the largest subpopulation of glial cells in the CNS) also express EPO following ischemic injury, and this response is known to ameliorate damage to the brain. To investigate the role of glial cells as a component of the systemic response to hypoxia, we created astrocyte-specific deletions of the murine genes encoding the hypoxia-inducible transcription factors HIF-1α and HIF-2α and their negative regulator von Hippel-Lindau (VHL) as well as astrocyte-specific deletion of the HIF target gene Vegf. We found that loss of the hypoxic response in astrocytes does not cause anemia in mice but is necessary for approximately 50% of the acute erythropoietic response to hypoxic stress. In accord with this, erythroid progenitor cells and reticulocytes were substantially reduced in number in mice lacking HIF function in astrocytes following hypoxic stress. Thus, we have demonstrated that the glial component of the CNS is an essential component of hypoxia-induced erythropoiesis. IntroductionThe erythropoietin (EPO) hormone is the chief regulator of red blood cell production in mammals, and its rate of synthesis is highly responsive to changes in physiologic oxygenation (1). Studies of this relationship between oxygen and red blood cell production date back to Viault's work at the end of the nineteenth century (2). Dissection of the mechanisms for induction of the Epo gene by low-oxygen conditions led to the discovery of a hypoxia responsive element (HRE) in the 3′ enhancer region of the gene; a binding site within the Epo gene was found for a family of transcription factors, which were subsequently termed hypoxia-inducible factors (HIFs) (3).HIF is a heterodimeric DNA-binding complex composed of 2 basic helix-loop-helix proteins of the PAS family: the constitutive non-oxygen-responsive subunit HIF-1β (also termed ARNT) and 1 of either hypoxia-inducible α-subunits, HIF-1α or HIF-2α (reviewed in refs. 4, 5). HIF-α subunits are rapidly degraded in normoxia but highly inducible by hypoxia. The interface between oxygen and the HIF-α subunits in normoxia is in part the hydroxylation of 2 prolyl residues in the oxygen-dependent degradation domain of the α subunits. HIF-α hydroxylation under normoxia regulates the interaction with the von Hippel-Lindau tumor suppressor protein (pVHL), which targets HIF-α for proteolysis by the ubiquitin-proteasome pathway (6).EPO synthesis is only detectable in a small number of tissues, including the kidney, liver, and the CNS (reviewed in refs. 7, 8). The kidney produces a large fraction of the circulating EPO in adult mammals (9), and it is clear that nonrenal EPO does not compensate for the loss of renal EPO production in patients with chronic

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MPScope: A versatile software suite for multiphoton microscopy

Cited by 99 publications

References 17 publications

Determination of scattering properties and damage thresholds in tissue using ultrafast laser ablation

Determination of scattering properties and damage thresholds in tissue using ultrafast laser ablation

Fluctuating and sensory-induced vasodynamics in rodent cortex extend arteriole capacity

The glial cell response is an essential component of hypoxia-induced erythropoiesis in mice

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