Summary: Analytical technique for studying substances present at enzyme concentrations in single cells, in situ, by measuring light absorption. Light from a tungsten strip lamp or xenon arc dispersed by a grating monochromator illuminates the optical system of a microscope. The absorbance of light is measured (in nanometers) by comparing the difference between the image of the sample and a reference image.

Top Publications

  1. Nymark S, Frederiksen R, Woodruff M, Cornwall M, Fain G. Bleaching of mouse rods: microspectrophotometry and suction-electrode recording. J Physiol. 2012;590:2353-64 pubmed publisher
    ..Our methods will now make possible an examination of bleaching adaptation and pigment regeneration in mouse animal lines with mutations or other alterations in the proteins of transduction. ..
  2. Ala Laurila P, Kolesnikov A, Crouch R, Tsina E, Shukolyukov S, Govardovskii V, et al. Visual cycle: Dependence of retinol production and removal on photoproduct decay and cell morphology. J Gen Physiol. 2006;128:153-69 pubmed
    ..The more rapid rates of retinol production in cones compared to rods are consistent with the more rapid operation of the visual cycle in these cells...
  3. Golobokova E, Govardovskii V. Late stages of visual pigment photolysis in situ: cones vs. rods. Vision Res. 2006;46:2287-97 pubmed
  4. Carleton K, H rosi F, Kocher T. Visual pigments of African cichlid fishes: evidence for ultraviolet vision from microspectrophotometry and DNA sequences. Vision Res. 2000;40:879-90 pubmed
    ..We have found evidence for ultraviolet visual capabilities in a Lake Malawi cichlid fish, Metriaclima zebra. Microspectrophotometry of single cones revealed a visual pigment with peak sensitivity at 368+/-4 nm. M...
  5. Govardovskii V, Fyhrquist N, Reuter T, Kuzmin D, Donner K. In search of the visual pigment template. Vis Neurosci. 2000;17:509-28 pubmed
    Absorbance spectra were recorded by microspectrophotometry from 39 different rod and cone types representing amphibians. reptiles, and fishes, with A1- or A2-based visual pigments and lambdamax ranging from 357 to 620 nm...
  6. Carleton K, Parry J, Bowmaker J, Hunt D, Seehausen O. Colour vision and speciation in Lake Victoria cichlids of the genus Pundamilia. Mol Ecol. 2005;14:4341-53 pubmed
    ..b>Microspectrophotometry determined that the wavelength of maximum absorption (lambdamax) of the rod pigment and three cone ..
  7. Parry J, Bowmaker J. Visual pigment coexpression in Guinea pig cones: a microspectrophotometric study. Invest Ophthalmol Vis Sci. 2002;43:1662-5 pubmed
    ..b>Microspectrophotometry was used to measure the absorbance spectrum of visual pigments in individual rods and cones from three ..
  8. Kindzelskii A, Sitrin R, Petty H. Cutting edge: optical microspectrophotometry supports the existence of gel phase lipid rafts at the lamellipodium of neutrophils: apparent role in calcium signaling. J Immunol. 2004;172:4681-5 pubmed
    ..Optical microspectrophotometry of Laurdan-labeled neutrophils revealed a large blue shift at lamellipodia relative to cell bodies...
  9. Rhee S, Miles E, Mozzarelli A, Davies D. Cryocrystallography and microspectrophotometry of a mutant (alpha D60N) tryptophan synthase alpha 2 beta 2 complex reveals allosteric roles of alpha Asp60. Biochemistry. 1998;37:10653-9 pubmed publisher
    ..These observations show that alpha Asp60 plays important roles in the closure of loop 6 and in allosteric communication between the alpha- and beta-subunits...

More Information


  1. Jain A, Pillai A, Sharma N, Verma K. Headspace single-drop microextraction and cuvetteless microspectrophotometry for the selective determination of free and total cyanide involving reaction with ninhydrin. Talanta. 2010;82:758-65 pubmed publisher
    ..used for the determination of cyanide with ninhydrin in combination with fibre-optic-based cuvetteless microspectrophotometry which accommodates sample volume of 1 microL placed between the two ends of optical fibres, and has been ..
  2. Parry J, Carleton K, Spady T, Carboo A, Hunt D, Bowmaker J. Mix and match color vision: tuning spectral sensitivity by differential opsin gene expression in Lake Malawi cichlids. Curr Biol. 2005;15:1734-9 pubmed
    ..By combining microspectrophotometry (MSP) of isolated cones, sequencing of opsin genes, and spectral analysis of recombinant pigments, we have ..
  3. Brown G, Chen D, Christianson J, Lee R, Stark W. Receptor demise from alteration of glycosylation site in Drosophila opsin: electrophysiology, microspectrophotometry, and electron microscopy. Vis Neurosci. 1994;11:619-28 pubmed
    ..We used electroretinography (ERG), microspectrophotometry (MSP), and electron microscopy (EM) to quantify visual cell defects...
  4. Sacré P, Deconinck E, Saerens L, De Beer T, Courselle P, Vancauwenberghe R, et al. Detection of counterfeit Viagra® by Raman microspectroscopy imaging and multivariate analysis. J Pharm Biomed Anal. 2011;56:454-61 pubmed publisher
    ..Application of the k-NN and SIMCA algorithm returned 100% correct classification during both internal and external validation. ..
  5. Emadi A, Wu H, de Graaf G, Enoksson P, Correia J, Wolffenbuttel R. Linear variable optical filter-based ultraviolet microspectrometer. Appl Opt. 2012;51:4308-15 pubmed publisher
    ..Experiments on a fabricated LVOF-based microspectrometer with this calibration approach implemented reveal a spectral resolution of 0.5 nm. ..
  6. Sano T, Suzuki S. Basic forensic identification of artificial leather for hit-and-run cases. Forensic Sci Int. 2009;192:e27-32 pubmed publisher
    ..was observed using scanning electron microscopy (SEM), color of these leather sheets was evaluated by microspectrophotometry (MSP), the leather components were measured by infrared micro spectrometry (micro-FT-IR) and the inorganic ..
  7. Taroni P, Valentini G, Comelli D, D Andrea C, Cubeddu R, Hu D, et al. Time-resolved microspectrofluorimetry and fluorescence lifetime imaging of hypericin in human retinal pigment epithelial cells. Photochem Photobiol. 2005;81:524-8 pubmed
    ..The latter change is observed also when high concentrations are reached locally, due to more efficient localization within the cell. ..
  8. Carleton K, Spady T, Streelman J, Kidd M, McFarland W, Loew E. Visual sensitivities tuned by heterochronic shifts in opsin gene expression. BMC Biol. 2008;6:22 pubmed publisher
    ..These resulting sensory shifts will have major impacts on visual communication and could help drive cichlid speciation. ..
  9. Barrett J, Siegel J, Goodpaster J. Forensic discrimination of dyed hair color: I. UV-visible microspectrophotometry. J Forensic Sci. 2010;55:323-33 pubmed publisher
    ..In this study, hair samples treated with 55 different red hair dyes were analyzed using UV-visible microspectrophotometry between 200 and 700 nm...
  10. Zhang Y, Wang Y. Distinct photopolymerization efficacy on dentin of self-etch adhesives. J Dent Res. 2012;91:795-9 pubmed publisher
    ..Raman spectral analysis disclosed that the chemical interaction between dentin and adhesives was responsible for the observations. We also verified this by tracking the distribution of the elements Ca and P in the adhesive layers. ..
  11. Vanderwee J, Porter G, Renshaw A, Bell M. The investigation of a relative contrast index model for fingerprint quantification. Forensic Sci Int. 2011;204:74-9 pubmed publisher
    ..Data from these separate sources will determine whether the theoretical aspects of the model would pragmatically produce reliable and repeatable results across a range of microspectrophotometers found in forensic laboratories. ..
  12. Jørgensen L, Schroeder T. Micro-lightguide spectrophotometry for tissue perfusion in ischemic limbs. J Vasc Surg. 2012;56:746-52 pubmed publisher
    ..O2C was easy to use, fast, and painless. The most useful finding was the high predictive value of a low saturation and the rise in O2C values after successful revascularization. ..
  13. Doukov T, Li H, Sharma A, Martell J, Soltis S, Silverman R, et al. Temperature-dependent spin crossover in neuronal nitric oxide synthase bound with the heme-coordinating thioether inhibitors. J Am Chem Soc. 2011;133:8326-34 pubmed publisher
    ..We found that some thioether inhibitors switch from high to low spin at lower temperatures similar to the "spin crossover" phenomenon observed in many transition metal complexes. ..
  14. Hart N, Lisney T, Marshall N, Collin S. Multiple cone visual pigments and the potential for trichromatic colour vision in two species of elasmobranch. J Exp Biol. 2004;207:4587-94 pubmed
    ..Using microspectrophotometry, we show that the retinae of the giant shovelnose ray (Rhinobatos typus) and the eastern shovelnose ray (..
  15. Arrese C, Hart N, Thomas N, Beazley L, Shand J. Trichromacy in Australian marsupials. Curr Biol. 2002;12:657-60 pubmed
    ..Using microspectrophotometry (MSP), we have investigated the spectral sensitivity of the photoreceptors of two Australian marsupials, ..
  16. Stavenga D, Stowe S, Siebke K, Zeil J, Arikawa K. Butterfly wing colours: scale beads make white pierid wings brighter. Proc Biol Sci. 2004;271:1577-84 pubmed
    ..The beads presumably cause the distinct matt-white colour of the wings of pierids and function to increase the reflectance amplitude. This will improve the visual discrimination between conspecific males and females. ..
  17. Rivera L, Blanco R, de la Villa P. Calcium-permeable glutamate receptors in horizontal cells of the mammalian retina. Vis Neurosci. 2001;18:995-1002 pubmed
    ..Fura-2 microspectrophotometry showed that in HCs, K+-induced cell depolarization promoted an increase in [Ca2+]i, mediated by the L-type ..
  18. Chan K, Kazarian S. New opportunities in micro- and macro-attenuated total reflection infrared spectroscopic imaging: spatial resolution and sampling versatility. Appl Spectrosc. 2003;57:381-9 pubmed
    ..One advantage of the diamond element is that it allows pressure to be applied and hence, good contact to be obtained over the whole field of view. ..
  19. Jokela Määttä M, Vartio A, Paulin L, Donner K. Individual variation in rod absorbance spectra correlated with opsin gene polymorphism in sand goby (Pomatoschistus minutus). J Exp Biol. 2009;212:3415-21 pubmed publisher
    ..microps. We propose that ambiguous selection pressures in the Baltic Sea and/or gene flow from the North Sea preserves polymorphism and is phenotypically evident as a wide variation in lambda(max). ..
  20. Nurizzo D, Cutruzzola F, Arese M, Bourgeois D, Brunori M, Cambillau C, et al. Conformational changes occurring upon reduction and NO binding in nitrite reductase from Pseudomonas aeruginosa. Biochemistry. 1998;37:13987-96 pubmed
  21. Sehić S, Betz G, Hadzidedić S, El Arini S, Leuenberger H. Investigation of intrinsic dissolution behavior of different carbamazepine samples. Int J Pharm. 2010;386:77-90 pubmed publisher
    ..Therefore, the intrinsic dissolution test with specific acceptance criteria can be a valuable and simple tool for monitoring, respectively reducing the variability of the CBZ bulk material. ..
  22. Marshall C, Olcott Marshall A. Raman hyperspectral imaging of microfossils: potential pitfalls. Astrobiology. 2013;13:920-31 pubmed publisher
  23. Didonna A, Vaccari L, Bek A, Legname G. Infrared microspectroscopy: a multiple-screening platform for investigating single-cell biochemical perturbations upon prion infection. ACS Chem Neurosci. 2011;2:160-74 pubmed publisher
  24. Moradian Oldak J, Du C, Falini G. On the formation of amelogenin microribbons. Eur J Oral Sci. 2006;114 Suppl 1:289-96; discussion 327-9, 382 pubmed
  25. Kavanagh K, Murphy E, Harmey M, Farrell M, Hardimann O, Perryman R, et al. Microspectrophotometric analysis of respiratory pigments using a novel fibre optic dip probe in microsamples. Physiol Meas. 1999;20:303-11 pubmed
    ..The key features of the system are small sample size, front-end adaptability, high sensitivity and fast multispectral acquisition which are essential for observing these biological reactions in vivo...
  26. Karan K, Yao X, Xu C, Wang Y. Chemical profile of the dentin substrate in non-carious cervical lesions. Dent Mater. 2009;25:1205-12 pubmed publisher
    ..Further studies are required to understand the role that these alterations play in response to acid etching and bonding to these clinically relevant substrates. ..
  27. Hoenicka H, Lautner S, Klingberg A, Koch G, El Sherif F, Lehnhardt D, et al. Influence of over-expression of the Flowering Promoting Factor 1 gene (FPF1) from Arabidopsis on wood formation in hybrid poplar (Populus tremula L. × P. tremuloides Michx.). Planta. 2012;235:359-73 pubmed publisher
    ..The latter observation was confirmed by UV microspectrophotometry on a cellular level...
  28. Jin M, Hwang S, Koo N, Kim B, Kho H, Choi S, et al. Autoantibodies in Sjögren's syndrome patients acutely inhibit muscarinic receptor function. Oral Dis. 2012;18:132-9 pubmed publisher
    ..SS IgG acutely and reversibly inhibited muscarinic receptor function, thereby inhibiting the Ca(2+) mobilization necessary for the activation of K(+) currents and ?-fodrin reorganization in HSG cells. ..
  29. Wang C, Li Y, Liu G, Ma J, Li Z, Wei X. Fiber confocal back-scattering micro-spectral analysis for single cell. Technol Cancer Res Treat. 2011;10:457-63 pubmed
    ..There was significant difference between the spectra of the normal and the cancer cells (p<0.001). This demonstrates that the FCBS system here is able to distinguish dysplastic cells from normal cells at cellular level. ..
  30. Parry J, Peirson S, Wilkens H, Bowmaker J. Multiple photopigments from the Mexican blind cavefish, Astyanax fasciatus: a microspectrophotometric study. Vision Res. 2003;43:31-41 pubmed
    ..of the expressed hypogean opsins of the hybrids, in comparison to the epigean forms, was undertaken by microspectrophotometry. The hybrid population showed considerable variation in the visual pigments of double cones, with evidence ..
  31. Gault N, Lefaix J. Infrared microspectroscopic characteristics of radiation-induced apoptosis in human lymphocytes. Radiat Res. 2003;160:238-50 pubmed
  32. Varela Álvarez E, Gómez Garreta A, Rull Lluch J, Salvador Soler N, Serrao E, Siguán M. Mediterranean species of Caulerpa are polyploid with smaller genomes in the invasive ones. PLoS ONE. 2012;7:e47728 pubmed publisher
    ..DAPI-staining with image analysis and microspectrophotometry were used to estimate relative nuclear DNA contents in three species of Caulerpa from the Mediterranean, ..
  33. Falcone P, Baiano A, Conte A, Mancini L, Tromba G, Zanini F, et al. Imaging techniques for the study of food microstructure: a review. Adv Food Nutr Res. 2006;51:205-63 pubmed
  34. GrasSo E, Yousef A, Rodriguez Romo L, Rodriguez Saona L. Rapid differentiation of Bacillus strains using hydrophobic grid membranes and attenuated total reflectance Infrared microspectroscopy. J Food Prot. 2009;72:1909-15 pubmed
  35. Bird B, Bedrossian K, Laver N, Miljkovic M, Romeo M, Diem M. Detection of breast micro-metastases in axillary lymph nodes by infrared micro-spectral imaging. Analyst. 2009;134:1067-76 pubmed publisher
    ..As a consequence, superior tissue classification and identification of cellular abnormality unattainable for deparaffinised tissue was achieved. ..
  36. Inokoshi M, Pongprueksa P, De Munck J, Zhang F, Vanmeensel K, Minakuchi S, et al. Influence of Light Irradiation Through Zirconia on the Degree of Conversion of Composite Cements. J Adhes Dent. 2016;18:161-71 pubmed publisher
    ..LI is too low through Ce-TZP/Al2O3 zirconia, necessitating the use of self-/dual-curing composite cements. ..
  37. O Neill T, Johnson J, Webster D, Gray C. The Canadian medicinal plant Heracleum maximum contains antimycobacterial diynes and furanocoumarins. J Ethnopharmacol. 2013;147:232-7 pubmed publisher
    ..This work supports the ethnopharmacological use of Heracleum maximum by Canadian First Nations and Native American communities as a treatment for infectious diseases, specifically tuberculosis. ..
  38. Mattana S, Alunni Cardinali M, Caponi S, Casagrande Pierantoni D, Corte L, Roscini L, et al. High-contrast Brillouin and Raman micro-spectroscopy for simultaneous mechanical and chemical investigation of microbial biofilms. Biophys Chem. 2017;229:123-129 pubmed publisher
    ..The correlative Raman and Brillouin investigation evidences the role of both extracellular polymeric substances and of hydration water in inducing a marked local softening of the biofilm. ..
  39. Feller K, Cronin T. Spectral absorption of visual pigments in stomatopod larval photoreceptors. J Comp Physiol A Neuroethol Sens Neural Behav Physiol. 2016;202:215-23 pubmed publisher
    ..Using microspectrophotometry (MSP) on larvae captured in the field, we found data which further support this long-standing hypothesis...
  40. Chen X, Liu X, Sheng D, Huang D, Li W, Wang X. Distinction of broken cellular wall Ganoderma lucidum spores and G. lucidum spores using FTIR microspectroscopy. Spectrochim Acta A Mol Biomol Spectrosc. 2012;97:667-72 pubmed publisher
    ..lucidum spores and G. lucidum spores. The result also indicates FTIR microspectroscopy may be useful for TCM identification. ..
  41. Ronda L, Bruno S, Bettati S, Mozzarelli A. Protein crystal microspectrophotometry. Biochim Biophys Acta. 2011;1814:734-41 pubmed publisher
    Single crystal microspectrophotometry has emerged as a valuable technique for monitoring molecular events that take place within protein crystals, thus tightly coupling structure to function...
  42. Ronda L, Bazhulina N, Morozova E, Revtovich S, Chekhov V, Nikulin A, et al. Exploring methionine ?-lyase structure-function relationship via microspectrophotometry and X-ray crystallography. Biochim Biophys Acta. 2011;1814:834-42 pubmed publisher
    ..enzyme towards L-methionine, substrate analogs and inhibitors was determined by polarized absorption microspectrophotometry. Spectral data were also collected for enzyme crystals, grown in monomethyl ether polyethylene glycol 2000 ..
  43. Yoshioka S, Matsuhana B, Tanaka S, Inouye Y, Oshima N, Kinoshita S. Mechanism of variable structural colour in the neon tetra: quantitative evaluation of the Venetian blind model. J R Soc Interface. 2011;8:56-66 pubmed publisher
    ..The experimental results and detailed analysis are found to quantitatively verify the model. ..
  44. Harding L, Qi S, Hill G, Reading M, Craig D. The development of microthermal analysis and photothermal microspectroscopy as novel approaches to drug-excipient compatibility studies. Int J Pharm. 2008;354:149-57 pubmed
    ..The study has therefore demonstrated the potential for using TAPM with PTMS for studying interactions at an individual particle level. ..
  45. Causin V, Schiavone S, Marigo A, Carresi P. Bayesian framework for the evaluation of fiber evidence in a double murder--a case report. Forensic Sci Int. 2004;141:159-70 pubmed
    ..Optical microscopy, UV-Vis microspectrophotometry and infrared analysis were employed to compare fibers recovered in the trunk of a car to those of the ..
  46. Barbour H, Archer M, Hart N, Thomas N, Dunlop S, Beazley L, et al. Retinal characteristics of the ornate dragon lizard, Ctenophorus ornatus. J Comp Neurol. 2002;450:334-44 pubmed
    The retina of a diurnal insectivorous lizard, Ctenophorus ornatus (Agamidae) was investigated using microspectrophotometry and light and electron microscopy. A prominent broad yellow band was observed that extended across the mid-retina...
  47. Grant S, Patel N, Philp A, Grey C, Lucas R, Foster R, et al. Rod photopigment deficits in albinos are specific to mammals and arise during retinal development. Vis Neurosci. 2001;18:245-51 pubmed
    ..The results strongly suggest that deficits in the rod cell population arise early in development of the mammalian albino retina, but do not generalize to nonmammalian mutants lacking retinal melanin. ..
  48. Lem J, Krasnoperova N, Calvert P, Kosaras B, Cameron D, Nicolo M, et al. Morphological, physiological, and biochemical changes in rhodopsin knockout mice. Proc Natl Acad Sci U S A. 1999;96:736-41 pubmed
  49. Kondrashev S. Long-wave sensitivity in the masked greenling (Hexagrammos octogrammus), a shallow-water marine fish. Vision Res. 2008;48:2269-74 pubmed publisher
    b>Microspectrophotometry (MSP) revealed that surprisingly for a "fully marine" species, in summer, photoreceptors of the nearshore scorpaeniform fish known as the masked greenling, Hexagrammos octogrammus, contained exclusively, ..
  50. Kapraun D. Nuclear DNA content estimates in green algal lineages: chlorophyta and streptophyta. Ann Bot. 2007;99:677-701 pubmed
    ..and red blood cell (chicken erythrocytes) standard were used to estimate 2C values with static microspectrophotometry. In Chlorophyta, including Chlorophyceae, Prasinophyceae, Trebouxiophyceae and Ulvophyceae, 2C DNA ..
  51. Sharma M, Crosbie J, Puskar L, Rogers P. Microbeam-irradiated tumour tissue possesses a different infrared absorbance profile compared to broad beam and sham-irradiated tissue. Int J Radiat Biol. 2013;89:79-87 pubmed publisher
  52. Lybeer B, Koch G, VAN Acker J, Goetghebeur P. Lignification and cell wall thickening in nodes of Phyllostachys viridiglaucescens and Phyllostachys nigra. Ann Bot. 2006;97:529-39 pubmed
    ..The lignification during ageing was studied topochemically by means of cellular UV microspectrophotometry. A combination of light microscopy and image analysis techniques were used to measure cell wall thickness...
  53. Pancaldi S, Baldisserotto C, Ferroni L, Bonora A, Fasulo M. Room temperature microspectrofluorimetry as a useful tool for studying the assembly of the PSII chlorophyll-protein complexes in single living cells of etiolated Euglena gracilis Klebs during the greening process. J Exp Bot. 2002;53:1753-63 pubmed
    ..These data suggest that microspectrofluorimetry may constitute a useful non-destructive tool for studying the assembly kinetics of PSII, under fully physiological life conditions...