Comparison of laser speckle and laser Doppler perfusion imaging: Measurement in human skin and rabbit articular tissue

Laser Doppler perfusion imaging (LDI) is currently used in a variety of clinical applications, however, LDI instruments produce images of low resolution and have long scan times. A new optical perfusion imager using a laser speckle measurement technique and its use for in vivo blood flow measurements are described. Measurements of human skin and surgically exposed rabbit tissue made using this instrument were compared with a commercial laser Doppler perfusion imaging instrument. Results from blood flow measurements showed that the laser speckle imager measured an 11–67% decrease in blood flow under arterial occlusion. Under similar conditions, the laser Doppler imager measured blood flow decreases of 21–63%. In comparison with LDI, it was observed that the higher temporal resolution of the laser speckle imager was more sensitive to measuring the hyperaemic response immediately following occlusion. This in vivo study demonstrated some of the several advantages laser speckle imaging has over conventional LDI, making the new instrument more versatile in a clinical environment.     

The effect of angiotensin II and vasopressin on renal haemodynamics

A comparison was made of the vascular actions of two hormones having a renal site of action, angiotensin II and vasopressin, using laser Doppler flowmetry to measure perfusion of the cortical and papillary regions of the kidney. Angiotensin II infusion caused dose-related increases in blood pressure and reductions in cortical perfusion, the latter responses being potentiated in the presence of the converting enzyme inhibitor, cilazapril. However, angiotensin II had no effect on papillary perfusion either before or following cilazapril. The reasons for this differing vasoconstrictor ability of angiotensin II at the cortex and papilla are unclear, but it could be due to medullary generation of prostaglandin or bradykinin. Administration of equipressor doses of vasopressin caused graded reductions in both cortical and papillary perfusions, and subsequent cilazapril significantly enhanced the papillary responses. This study demonstrates that the regulation of blood flow through the different regions of the kidney can be differentially regulated by the peptide hormones angiotensin II and vasopressin.     

Integrating probe for tissue laser Doppler flowmeters

Methods to measure microvascular blood flow usually probe small volumes of tissue. Therefore, spatial differences in skin blood flow alter the signal, when the sensing element is moved a short distance. To reduce the effects of spatial differences in skin blood flow, but yet record its temporal variability, a new integrating probe for laser Doppler flowmeters was developed. The probe receives light from seven different scattering volumes simultaneously, and the instrument processes an integrated signal which is ultimately taken as the average flow value. Significant spatial integration is found, as spatial variability is reduced by the square root of the number of scattering volumes.     

Oxford Optronix MPM 3S: a clinical assessment of a microvascular perfusion monitor.

The Oxford Optronix MPM 3S is a new microvascular perfusion monitor which is promoted as a device for use in the operating theatre. It uses a semiconductor laser diode and applies the Doppler principle to derive a semi-quantitative estimation of microvascular flow. We assessed this instrument with eight healthy volunteers who each performed eight different orthostatic arm manoeuvres while forearm skin blood flow was monitored. The different manoeuvres caused statistically significant changes in the instrument's reading which generally were consistent with expected changes in blood flow. The monitor also was assessed in the theatre environment with four anaesthetized patients. It proved easy to use, and was not subject to electrical interference from other equipment including short-wave diathermy. The major practical limitation of the technique is the semi-quantitative nature of the measurement. The instrument appears to have potential clinical uses in plastic and vascular surgery.     

Effects of alternating magnetic fields and low-frequency electric currents on human skin blood flow

The influence of alternating magnetic fields on human skin blood flow was investigated. The hands of volunteers were fixed just above an induction heater and the regional skin blood flow of a fingertip was measured by a laser Doppler flowmeter. The hands were exposed to fields with flux densities of 16 mT, 32 mT and 48 mT at a frequency of 3·8 kHz. Two parameters of the laser Doppler blood flow recording were studied: the dynamic and DC components. The dynamic component did not show any particular changes following magnetic stimulation. The DC component showed a rapid decrease in blood flow in the first period after field-on at 48 mT and 32 mT. A pain sensation with muscle contraction was recognised (48 mT and 32 mT). The effect of electrical stimulation on skin blood flow was also investigated. The differences between the magnetic and electrical effects are discussed. The results suggest that the human body responds to magnetic stimulation with an escape reaction which results in an elevated vasoconstrictor tone. The change in microvascular blood flow recorded with the laser Doppler method can probably be explained by the influence of the eddy currents on the sensory receptors of the skin.     

Comparative study of two laser Doppler blood flowmeters

A new infrared laser Doppler blood flow instrument (Moor MBF3D) was evaluated using an in vitro model allowing measurements over a range of flow velocities and concentrations. The responses correlated well (r = 0.96, p less than 0.01) with those obtained simultaneously using a Perimed PF3 laser Doppler instrument. The different processing bandwidths of the instruments were investigated and the wideband mode of operation is recommended for flow measurements where there may be fast moving red blood cells (rbcs). The infrared instrument is capable of dual-channel operation, and the two channels are shown to respond almost identically for similar changes in blood flow through the in vitro model (r = 0.999, p less than 0.01). The main advantage of the dual-channel instrument is that continuous measurements may be made simultaneously at two different skin sites allowing dynamic flow responses to be compared.     

Different responses of Finapres and the oscillometric finger blood pressure monitor during intensive vasomotion

Two different techniques for non-invasive beat-to-beat finger arterial blood pressure monitoring are compared in six healthy volunteers during local hand heating from 21 to 38^°C. The degree of peripheral vasoconstriction was established by recording the thumb pulp skin blood flow with a laser Doppler instrument. For time episodes without vasoconstriction no systematic difference in the readings of beat-to-beat mean blood pressure of the two monitors was found (the oscillometric device UT9201 minus Finapres differencewas 0.3 mm Hg, SD 0.3). For the episodes with vasoconstriction the difference was statistically significant (6.7 mm Hg, SD 2.0). The oscillometric device minus Finapres difference and the laser Doppler signal were found to be inversely correlated, the correlation coefficient varying from - 0.28 to - 0.67. A disagreement between the readings of the instruments during intensive vasomotion is assumed to be caused mainly by the tendency of the oscillometric method to overestimate the finger mean blood pressure under the condition of peripheral vasoconstriction.     

Effect of inhibition of peptidase activity on distribution of intrarenal blood flow.

Experiments were performed in dogs to determine the effects of the intravenous administration of the dipeptide hydrolase inhibitor SQ 20,881 on renal hemodynamics, intrarenal blood flow distribution, and renal function. Dipeptide hydrolase converts angiotensin I to angiotensin II and inactivates bradykinin. SQ 20,881 causes an inhibition of the vasoconstrictor response after angiotensin I and potentiation of the vasodilatory activity of bradykinin. Total renal blood flow, cortical distribution of blood flow, and glomerular filtration rate were determined. In seven animals administration of SQ 20,881 (1 mg/kg) resulted in a decrease in mean systemic blood pressure of 11 mmHg, an increase in total renal blood flow of 0.71 ml/min per g, and a significant fall in glomerular filtration rate. Fractional blood flow to the superficial cortex decreased and to the juxtamedullary cortex increased. Absolute flow was unchanged in the superficial cortex and increased significantly in the deep cortex. The findings are compatible with reported effects of bradykinin on intrarenal blood flow distribution, although the experiments do not distinguish between potentiation of bradykinin or inhibition of angiotensin I conversion.     

Heterogeneous blood flow response in the foot on dependency,assessed by laser Doppler perfusion imaging

The exact nature of the decrease in foot skin blood flow seen after a change in posture remains unsettled. This mechanism has previously been examined by non-invasive techniques such as the laser Doppler perfusion monitor (laser Doppler flowmetry). Taking into account the shortcomings of laser Doppler perfusion monitoring when applied to the determination of skin blood flow, which normally shows substantial heterogeneity, we have applied an emerging technology, the laser Doppler perfusion imager (LDPI). This technique provides a more comprehensive picture of the blood flow distribution in the skin, as it maps skin blood flow over a surface area (120×120 mm, 4096 measurement sites). It was used to examine if the reduction in tissue perfusion or the alterations in flow distributions seen after a change in posture (supine to dependency) could be fully explained by an increase in venous pressure (venous stasis of 50 mmHg) or if the data suggest a complementary mechanism. Skin blood flow of the forefoot decreased from 0.60 V (volt) (median) during rest to 0.40 and 0.38 V during venous stasis and dependency, respectively. Although almost identical median values were obtained during stasis and dependency, the flow distributions were different, with a loss of high flow values during venous stasis. Biological zero was 0.24 V. As the LDPI technique readily records skin perfusion during variations in venous stasis and posture, as well as information on flow distribution changes, it appears promising for future application in stimuli-response studies of skin blood flow. The difference in flow distribution seen between increased venous pressure and dependency suggests an additive regulatory mechanism to the veni-vasomotor reflex during a change in posture.     

Localized neural control of blood flow in the renal cortex of the anaesthetized baboon

Localized neural control of renal cortical blood flow was observed in five chloralose-anaesthetized baboons. Renal artery blood flow was measured with an electromagnetic flowmeter, while superficial renal cortical blood flow was measured with a laser Doppler flowmeter. Stimulation of all renal nerve bundles isolated produced significant reductions in total renal blood flow, but reduced the superficial renal cortical blood flow only in localized areas. Systemic arterial blood pressure did not vary significantly, which suggests that the changes in renal cortical blood flow were neurally mediated and not the result of autoregulation. The renal cortical blood flow response was delayed 1-4 s with respect to the total renal blood flow response. The localization of flow control is consistent with the neuroanatomy of the kidney, and the delayed renal cortical blood flow response is consistent with the vascular anatomy of the kidney and with previous measurements of red cell transit times through the renal cortex.     

Prostaglandin E2 is required for ultraviolet B-induced skin inflammation via EP2 and EP4 receptors

Keratinocytes are the major target of sunlight, and they produce prostaglandin (PG) E(2) upon ultraviolet (UV) exposure. Although indomethacin, one of cyclooxygenase inhibitors, is known to suppress UV-induced acute skin inflammation, it remains uncertain whether endogenous PGE(2) is responsible for UV-induced skin inflammation, and which subtype of PGE(2) receptors mediates this process. UV-induced skin inflammation was investigated by using genetically and pharmacologically PGE(2) receptor-deficient mice. We applied UV-induced skin inflammation model to genetical and pharmacological PGE(2) receptor-deficient mice. We exposed UVB on these mice at 5 kJ/m(2), and examined the ear swelling and the histological findings. We also measured the blood flow using a laser doppler device to assess the intensity of UVB-induced inflammatory change. The UV-induced ear swelling at 48 h after exposure was significantly reduced in EP2(-/-), EP4(-/-) or wild-type mice treated with the EP4 antagonist compared to control mice. Consistently, inflammatory cell infiltration into the local skin, and local blood flow after UV exposure were significantly reduced by EP2 or EP4 signaling blockade. These data suggest that PGE(2)-EP2/EP4 signaling is mandatory in UV-induced acute skin inflammation, presumably by enhancing blood flow in the microenvironment.     

Similarity of fingertip skin blood flow patterns recorded by the model-based thermal clearance and large area laser Doppler probes.

We compared two different techniques for non-invasive registration of fingertip skin blood flow patterns in 15 healthy volunteers during a rest at room temperature 26-28 degrees C. The large area probes of a two-channel laser Doppler flowmeter (LDF) and an experimental probe of a thermal clearance (TC) instrument provided approximately the same sample volume of the studied tissue. The laser Doppler probes were attached to the pulp of the index (LDF1) and ring (LDF2) fingers, while the TC probe to the pulp of the middle finger. A computer simulation method, provided by the MATLAB package, was introduced to speed up the frequency response of the TC probe. By calculating the correlation coefficients and applying the least squares criterion, we evaluated the similarity of the 3-min samples having time resolution of 1 s. The group-averaged value of the correlation coefficient (median with a 99% confidence interval) for LDF2 versus LDF1 equalled 0.95 (0.92-0.97). After an introduction of the computer-based frequency correction, the correlation coefficient for TC versus LDF1 increased from 0.81 (0.61-0.85) till 0.92 (0.84-0.95), while the root mean squared error between TC and LDF1 diminished by 33% (p<0.001).     

Continuous mean arterial pressure measurement in the fingers: The influence of local arm cooling

Two different methods were studied for non-invasive beat-to-beat finger mean arterial pressure (MAP) measurement in 14 healthy volunteers during local arm cooling to 17°C. Fingertip skin blood flow was recorded by a laser Doppler instrument. The difference between the oscillometric beat-to-beat blood pressure measurement device (UT9201) and volume clamp monitor (Finapres) before cooling was −1.5±1.1 mmHg, and, although statistically significant, (p<0.01), this difference cannot be considered clinically meaningful. The results obtained using the two devices during arm cooling showed a systematic disagreement in the continuously recorded MAP. After a 30s period of cooling, the group-averaged difference of UT9201 minus Finapres, calculated as change from the baseline, reached (mean±SD) 8.8±6.3 mmHg (statistically significant, p<0.01).     

The distribution of renin-containing cells in kidneys with renal artery stenosis—an immunocytochemical study

In 10 kidneys removed for hypertension due to renal artery stenosis, the histological appearances varied from negligible ischaemic damage to end-stage ischaemic atrophy. We stained the renin-containing cells in tissue sections using an antiserum to pure human renin and an immunoperoxidase technique. In all kidneys there was hyperplasia of the renin-containing cells both in juxtaglomerular apparatuses (JGAs) and in arteries outside the JGA, where these cells extended proximally as far as the Interlobular arteries. We mapped the distribution of renin-containing cells and found them in all zones of the renal cortex; in three kidneys they were predominantly in the superficial cortex; in four they were distributed more evenly throughout the width of the cortex; but in three kidneys the normal gradient was reversed, with most of the cells being in the juxtamedullary cortex and, in two of the cases, little or no stainable renin in the superficial cortical JGAs. We suggest that these abnormalities in the distribution of renin-containing cells could affect both the pattern of intrarenal blood flow and the site in the kidney at which secreted renin enters the blood.     

Physical assessment of medical ultrasound instruments

The Oxford Optronix MPM 3S is a new microvascular perfusion monitor which is promoted as a device for use in the operating theatre. It uses a semiconductor laser diode and applies the Doppler principle to derive a semi-quantitative estimation of microvascular flow. We assessed this instrument with eight healthy volunteers who each performed eight different orlhostalic arm manoeuvres while forearm skin blood flow was monitored. The different manoeuvres caused statistically significant changes in the instrument's reading which generally were consistent with expected changes in blood flow. The monitor also was assessed in the theatre environment with four anaesthetized patients. It proved easy to use, and was not subject to electrical interference from other equipment including short-wave diathermy. The major practical limitation of the technique is the semi-quantitative nature of the measurement. The instrument appears to have potential clinical uses in plastic and vascular surgery.     

Acute exercise causes an enhancement of tissue renin–angiotensin system in the kidney in rats

Aims: Initially, the renin–angiotensin system (RAS) produced through the classical endocrine pathway was well known for its regulation of blood pressure. However, it was revealed that a local autocrine and/or paracrine RAS may exist in a number of tissues (such as kidney). Exercise causes a redistribution of tissue blood flow, by which the blood flow is greatly increased in active muscles, whereas it is decreased in the splanchnic circulation (such as in the kidney). We hypothesized that exercise causes an enhancement of tissue RAS in the kidney. Methods: We studied whether exercise affects expression of angiotensinogen and angiotensin‐converting enzyme (ACE) and tissue angiotensin II level in the kidney. The rats performed treadmill running for 30‐min. Immediately after this exercise, kidney was quickly removed. Control rats remained at rest during this 30‐min period. Results: The expression of angiotensinogen mRNA in the kidney was markedly higher in the exercise rats than in the control rats. ACE mRNA in the kidney was significantly higher in the exercise rats than in the control rats. Western blot analysis confirmed significant upregulation of ACE protein in the kidney after exercise. Tissue angiotensin II level was also increased by exercise. Conclusion: The present study suggests that the exercise‐induced enhancement of tissue RAS in the kidney causes vasoconstriction and hence decreases blood flow in the kidney, which are helpful in increasing blood flow in active muscles, thereby contributing to the redistribution of blood flow during exercise.     

Laser-Doppler flowmetry

Laser-Doppler flowmetry is a new method of the continuous and noninvasive measuring of the tissue blood flow, utilizing the Doppler shift of laser light as the information carrier. The method has already proved its potential usefulness in the clinical assessment of blood flow within disciplines such as dermatology, plastic surgery, and gastrointestinal surgery. In experimental medicine, laser-Doppler flowmetry has been used in the study of spontaneous rhythmical variations as well as in the study of spatial and temporal fluctuations in human skin blood flow. The method has facilitated further investigations of the nature of blood flow in a variety of tissue types. The method is rapidly spreading to many new applications in microvascular blood flow measurements. Several commercial laser Doppler instruments are available. This review article is a presentation of the state of the art of laser-Doppler flowmetry including theory, technology, and applications.     

Measurement of the skin microcirculation through intact bandages using laser Doppler flowmetry

The microcirculation under compression bandages has been assessed by numerous methods; however, the measurement techniques can disrupt the bandage-skin interface, affecting the measurement. In this study, a non-invasive method for measuring cutaneous blood flow using laser Doppler flowmetry (LDF) is presented. Ten volunteers had their microcirculation assessed by a laser Doppler probe being placed on their upper forearm with and without a light-transmissive gel and with a compression bandage plus light-transmissive gel. A circulatory challenge to the bandaged forearm in two of the volunteers was also undertaken. The median (95% confidence interval) perfusion (p.u.) for the skin surface was 24 (15–33) perfusion units (p.u.), and the skin plus light-transmissive gel demonstrated a higher perfusion: 66 (50–82) p.u., (p<0.012). The addition of the compression bandage, with additional gel allowed to permeate through to the underlying skin, decreased the perfusion to 27 (20–34) p.u. (p<0.007). In both volunteers, the microcirculatory flow responded to the vascular challenge, resulting in flow changes related to the cuff pressure (45-27 and 14-8 p.u.). This method demonstrated that it may be possible to assess the microcirculation through intact bandages, without the need to place any sensors at the skin-bandage interface.     

Analysis of fluctuations of capillary blood flow in the cortex of rat kidneys during ischemia

The dynamics of fluctuations of capillary blood flow in the cortex of rat kidneys under conditions of transient occlusion of renal arteries and veins or simultaneous occlusion of afferent and efferent vessels was studied by laser Doppler flowmetry. The initial variability of blood flow in the right kidney was higher than in the left kidney. The most pronounced changes in the right kidney were observed during occlusion and subsequent recirculation. In the left kidney, the most pronounced changes were found during occlusion and after arterial occlusion. Spectral analysis gave similar results. The recovery of renal blood flow after transient venous occlusion took longer time than after arterial occlusion (especially in the right kidney). Translated fromByulleten' Eksperimental'noi Biologii i Meditsiny, Vol. 128, No. 8, pp. 148–152, August, 1999     

Mechanism of intrarenal blood flow redistribution after carotid artery occlusion.

Bilateral carotid artery occlusion results in an increase in mean arterial pressure, an increase in renal sympathetic nerve activity, and a redistribution of renal blood flow from inner to outer cortex. To elucidate the mechanism of the renal blood flow redistribution, carotid artery occlusion was performed in anesthetized dogs with the left kidney either having renal perfusion pressure maintained constant (aortic constriction) or having alpha-adrenergic receptor blockade (phenoxybenzamine); the right kidney of the same dog served to document the normal response. When renal perfusion pressure was maintained constant, renal blood flow distribution (microspheres) was unchanged by carotid artery occlusion. In the presence of renal alpha-adrenergic receptor blockade, carotid artery occlusion elicited the usual redistribution of renal blood flow from inner to outer cortex. The redistribution of renal blood flow observed after carotid artery occlusion is mediated by the increase in renal perfusion pressure rather than the increase in renal sympathetic nerve activity.