The document discusses the Nobel Prize in Chemistry, particularly the 2017 award to Joachim Frank for his development of cryo-electron microscopy, a technique vital for imaging biomolecules. It outlines the principles of cryo-electron microscopy, its applications in structural biology, and the methodology involved in preparing samples for imaging. Additionally, it highlights the advantages and future prospects of cryo-em technology in various scientific fields, including pharmacology and drug design.

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2. Nobel Prizein Chemistry2017
JoachimFrank
Rabia Aziz
BS-IV Organic Chemistry
Jinnah University for Women
Karachi
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3. Introduction

4. Nobel Prizein Chemistry
• The Nobel Prize in Chemistry is an awarded annually by the Royal Swedish
Academy of Sciences to scientists.
• It is one of the five Nobel Prizes established by the will of Alfred Nobel in 1895,
awarded for outstanding contributions in chemistry, physics, literature, peace,
and physiology or medicine.
• The award is presented in Stockholm at an annual ceremony on December 10,
the anniversary of Nobel's death.
• From 1901 to 2017, the award has been bestowed on a total of 177 individuals.
• A Chemistry Nobel Prize laureate earns a gold medal, a diploma bearing a
citation, and a sum of money
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6. Joachim Frank, Ph.D.
Profession:
Biography:
Fields of Specialization:
(1) Application of electron microscopy and image
analysis to the elucidation of three-dimensional
structure and function of macromolecular
assemblies and cell components.
(2) The mechanism of protein synthesis
2017NobelPrize Winner
Nobel Prize:
He was honoured for his
development of cryo-electron
microscopy, which improves the
imaging of biomolecules.11/7/2018 6Rabia Aziz
JoachimFrank’s

7. Thesis
1. Investigation of secondary electron
emission of gold at its melting point
2. Investigations of high-resolution
electron micrographs using image
difference and reconstruction methods
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1. Theory of partial coherence in electron
microscopy
2. Single-particle approaches in electron
microscopy
Working
1. Found in Translation –
Collection of Original
Articles on Single-Particle
Reconstruction and the
Structural Basis of Protein
Synthesis
2. Molecular Machines in
Biology: Workshop of the
Cell
3. Electron Crystallography of
Biological Macromolecules
4. Electron Tomography (2nd
ed.)
5. Three-Dimensional
Electron Microscopy of
Macromolecular
Assemblies (2nd ed.)
Books
Articles
1. "Generalized single-particle cryo-EM – a historical
perspective".
2. "Structure and dynamics of a processive Brownian
motor: the translating ribosome".
3. "The ribosome and the mechanism of protein
synthesis".
4. "Single-particle reconstruction of biological
macromolecules in electron microscopy -- 30 years"

8. Techniques for Bio-moleculesAnalysis
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9. Cryo-ElectronMicroscopy

10. • Classification:

11. Electron cryomicroscopy
(CryoEM)
is an electron microscopy (EM) technique
where the sample is cooled to cryogenic
temperatures.
 Transmission electron cryomicroscopy
• Transmission electron cryomicroscopy
(CryoTEM) is a transmission electron
microscopy technique that is used in
structural biology.
• Electron cryotomography (CryoET), a
specialized application of where samples are
imaged as they are tilted
• Single particle CryoEM
• In single-particle reconstruction, the image of
each complex is formed.
 Scanning electron cryomicroscopy
• Scanning electron cryomicroscopy (CryoSEM),
is scanning electron microscopy technique
with a scanning electron microscope's cold
stage in a cryogenic chamber.

12. Cryo-ElectronmicroscopyCharacteristics
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1. Native state of specimen; not stained not
Fixed
2. Specimens are observed in vitreous ice
3. Vitreousbe maintained in microscope ice is
water frozen to -140° C in less than 10-4 sec
4, Low dose parameters are required so the
sample is not destroyed
5. Automated 3D image to get high resolution
images
Cryo-electron microscopy (cryo-EM) and cryo-electron
tomography are employed in structural biology to determine
the structure of vitrified biological samples, such as isolated
proteins and protein complexes, two-dimensional (2D)
protein/lipid crystals, or larger biological objects such as
bacteria and nanocrystals.

14. Cryo-ElectronMicroscopyProcess
Take a sample
Cool it so rapidly that it is
immobilized before it has
time to crystallize
Cut it into ultra-thin
sections
Observe, in a cryo-
electron microscope
Reconstruct the 3-D
distribution of the
material in the section by
means of computerized
electron tomography CET
The time span from the sample
to the image could be a as say
short as one hour rather than
day
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Preparation of the material
Vitrification
Cryo-sectioning
Observing in the cryo-electron microscope
Cryo-EM is a method that uncovers the structure of
macromolecular complexes. It involves purifying a
solution of a certain macromolecular complex,
placing the solution onto a grid, freezing it
(cryogenically), and then imaging the frozen film
using an electron microscope. This produces many
images of the same complex in different
orientations.

15. SampleAnalysis
Grid preparation method Typical samples Typical results
Cryogenic visualisation Macromolecular complexes
(>150 kDa, liposomes,
organelles, prokaryotic and
eukaryotic cells
Visualisation of specimen
with low contrast
Cryogenic reconstruction
(single particle)
Macromolecular complexes
(>150 kDa
Ribosomes, Icosahedral
Viruses
Depending on the specimen,
reconstructions of ∼3–20 Å
Cryogenic reconstruction
(tomography)
Organelles, prokaryotic cells,
thin edge or lamellar of
eukaryotic cells
Resolution of
tomograms < 10 nm,
subtomogram averaging can
produce reconstructions of
>10 Å

16. Instrumentation
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17. Resolution
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Using cryo‐EM we can now calculate
near‐atomic resolution structures from
relatively few thousand images of
macromolecular complexes with sizes
ranging from ∼150 kDa to several MDa.
Typical usage: A wide range of samples, from
small (∼150 kDa) macromolecules to whole
eukaryotic cells can be vitrified and imaged by
cryo-EM.

18. Imagingand DataProcessing
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Computer Data Processing

19. Joachim Frank Image Analysis

20. FrankRefinesImageAnalysis:
• In 1975, Joachim Frank began work on the algorithms that would
analyze fuzzy 2D images and reconstruct them into sharp 3D
structures.
• He developed a mathematical method that allowed the computer
to identify different recurring patterns in the image.
• The computer then sorted similar patterns into the same group
and merged the information in these images to generate an
averaged, sharper image. In this way he obtained a number of
high-resolution, two-dimensional images that showed the same
protein but from different angles. The algorithms for the software
were complete in 1981.
• Joachim Frank develops Spider software for conversion of
2D images into 3D structures.

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cryo‐sample preparation, data collection and processing workflows, and the
modeling and refinement of structures in cryo‐EM 3D maps

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Figure 1. Main conceptual steps of processing.
SPIDER Reconstruction Engine, or SPIRE.

23. Cryo-EM
Relationships
with
Chemistry
Biochemistry
Microbiology
Pharmacy
Computer
Science
Physics
Biotechnology
Nanotechnolo
gies

24. Chemistry
• The structure of ribosomes is interesting in the context of antibiotic
development since many antibiotics target the bacterial ribosome.
Cryo-EM is revolutionising the whole
field of structural biology and will be
particularly important in cancer drug
design.

25. Advantagesand Disadvantages
Advantages:
• Sample is visualised in a native-like state. Processing of
data can yield high-resolution, 3D structural information.
• A great advantage with respect to pharmacological
application is the possibility to identify with high precision
the single amino acids residues and their ligands, as well
as hydrogen bonds and intermolecular interactions.
Disadvantages:
• Biological samples must be imaged with low electron
doses to prevent radiation damage resulting in poor
contrast.
• It is sometimes non-trivial to optimise sample distribution
and ice thickness.

26. Applications
1. Nanoparticle Research
2. Pharmaceutical Drug Research
3. 3D Structure Visualization of: Single Particles
such as Ribosome, tRNA, Viruses, Proteins,
Macromolecules; Lipid Vesicles
4. Structure and dynamics study
Cryo-EM has a huge potential for the
clarification of pathological mechanisms
and it could assist the early identification
and selection of drug candidates and the
validation of pathological targets.
Virology, neurosciences, tumors and
immunology are the most promising
areas of application,

27. FutureProspects:
• A number of improvements are expected in future
• High Voltage Electron Source
• Mathematical Correction of Lens Defects
• High Resolution
• Improved Scanners
• Better High-Pressure Freezing
• Improved CCD detectors will remove the need for
computer processing in future
Advancement: A hybrid approach involving the combination of structure
prediction, prospectrometry (CX‐MS), and identification and modeling of the
corresponding segment in the high‐resolution cryo‐EM map.

28. Conclusion:
• Electron cryomicroscopy (CryoEM) is an electron
microscopy (EM) technique where the sample is cooled to
cryogenic temperatures.
• The cryo-EM method can be used to determine the three-
dimensional structure of biomacromolecules in near native
condition at close to atomic resolution, and has the
potential to reveal conformations of dynamic molecular
complexes.
• Joachim Frank made the technology generally applicable.
Between 1975 and 1986 he developed an image processing
method in which the electron microscope’s fuzzy two
dimensional images are analysed and merged to reveal a
sharp three-dimensional structure.

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