Raman spectroscopy is highly sensitive in the detection of haem proteins and is ideal for studying their redox biology, something that can be achieved without requiring these proteins to be isolated or stained. The technique can be used to determine redox states within biological systems, allowing scientists to research redox dynamics and the effects they have on health regulation and disease.
The redox biology of haem proteins is closely related to their functions which include oxygen transport, oxygen storage, electron transport and free radical scavenging. The images produced by Raman spectra enable visualization of the spatial relationships, distribution, redox and spin states of haem proteins within biological systems. These properties can also be compared between samples and in-depth data can be gained about the biological system being studied. The data generated can be processed to provide a wide range of information and examples of this are described below.
Haem protein type and electronic states
Characteristic Raman spectra can be used to identify haem protein type and the Raman band positions of these proteins can be used to determine their redox and spin states. Using resonance Raman spectroscopy with visible wavelengths, the signal intensities of these protein bands can be specifically enhanced, thereby providing high sensitivity. Raman imaging can be directly performed on haem proteins within live/fixed cellsi, whole organsii or sections of tissueiii.
Haem protein within a biological system
Using Raman spectroscopy, detailed, sub-micrometer spatial information can be obtained and the exact location of haem proteins within a sample can be clearly shown. Information on the relationship between their distribution, protein, organelle and cell functions can also be obtained.
Using redox biology to understand protein, organelle and cell functions and dynamics
Raman imaging can provide insight on the correlation between the redox state of cytochromes with mitochondrial intermembrane potential and energy productioni. The technique can help researchers better understand the relationship between mitochondrial dysfunction and conditions such as infertility or neurodegenerative diseases.
The relationship between the distribution of hemoglobin in erythrocytes and the oxygen transport functions/abnormalities of haem proteins can also be investigated. Other systems the technique can shed light on include synthetic biology and tissue engineering, such as the oxidoreductase function of man-made redox proteinsiv and oxygenation function of myoglobinv.
Raman imaging as a tool for biological research
Raman spectroscopy is ideal for use in redox biology research, enabling comparison of physical and electronic properties of haem proteins between samples. Chemical information on other biomolecules including lipids, proteins, carbohydrates, nucleic acid and minerals can all be obtained simultaneouslyiii and valuable insights can be gained into the effects redox regulation has on biological systems.
Renishaw’s inVia confocal Raman microscope
This research-grade confocal Raman microscope from Renishaw features SreamLineTM imaging technology that allows for high-speed mapping of haem proteins, without causing any damage to cells or tissues. The option of StreamLine imaging with Slalom allows researchers to quickly gain an overview of tissue samples, while high-confocality SreamLineTM imaging is available for the scrutiny of small details.
Watch our webinar about Resonance Raman spectroscopy for redox biology research.
i. Brazhe et al (2012) PLoS ONE 7(9): e41990
ii. Brazhe et al (2013) PLoS ONE 8(8): e70488
iii. Ramser et al (2012) Applied Spectrosc 12: 1454-60
iv. Anderson et al (2014) chem Sci 5: 507-514
v. Armstrong et al (2015) Nature Comm 6: 7405
Renishaw is a global company and a recognised leader in Raman spectroscopy. Offering high performance optical spectroscopy products for over 20 years, Renishaw’s Spectroscopy Division is passionate about manufacturing the very best Raman products. It has a team of scientists and engineers specialising in the development, application and production of high performance, configurable Raman spectrometers.
With a foundation built on sound engineering, these systems offer the highest levels of flexibility and performance and are used by academics and industrialists to tackle analytical problems across a broad range of application areas, including life science, chemicals; materials science; pharmaceuticals; semiconductors; forensics; gemmology; antiquities; and green energy, such as photovoltaics.
Renishaw’s Raman systems are easy to use and produce repeatable, reliable data, even from challenging samples. We aim to be a long-term partner, offering quality products that meet customers’ needs, both today and into the future, backed up by expert technical and commercial support.
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