Estimation and Imaging Techniques to Study Lipids in Mammalian Samples

  • Sudhanshu Shukla
  • Sanghamitra Mishra
Part of the Springer Protocols Handbooks book series (SPH)


Lipids are small biomolecules that constitute structural elements of the cellular membranes and play crucial functions to maintain various cellular processes indispensable for survival. In general, there are four types of lipids existing in the human body mainly fatty acids, glycerides, non-glycerides, and complex lipids. Abnormal lipid levels and signaling give rise to a number of clinical pathologies and disorders including metabolic diseases, cardiovascular diseases, diabetes, and immune dysfunctions. Some of these, such as sitosterolemia, hypercholesterolemia, fish eye disease, age-related macular degeneration, Tangier disease, etc., are inherited and caused by mutations in genes from lipid pathways. Therefore, it is important to study the molecular mechanisms of disease progression and establishing correlation between lipid levels and grade of clinical onset to understand the pathophysiologies of various lipid-associated diseases. This requires reliable imaging and quantification techniques of various lipids. In this chapter, by taking samples from human eyes and cell culture models, we have explained how imaging and quantification techniques can provide molecular details toward understanding the correlation between lipids and eye disease. We describe the use of standard kits and techniques available, and some of these protocols can be extrapolated to other organ systems with specific modifications.


Membrane lipids Cholesterol LDL FM dyes Filipin 



The images shown in this chapter were part of work done in Dr. Graeme J. Wistow’s lab, supported by Intramural Program of the National Eye Institute. We thank Dr. Graeme J. Wistow, Dr. Katherine Peterson, Dr. Dinusha Rajapakse, Dr. Campos Maria, and Biological Imaging Core of the National Eye Institute Bethesda for kindly sharing the images.


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© Springer Science+Business Media, LLC, part of Springer Nature 2020

Authors and Affiliations

  • Sudhanshu Shukla
    • 1
  • Sanghamitra Mishra
    • 2
  1. 1.School of MedicineCase Western Reserve UniversityClevelandUSA
  2. 2.MedGenome Labs Ltd.BangaloreIndia

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