Download pdf - Interpretation of SEM images-iolo-printout images.pdfRomanesque Brocolli/Cauliflower (or Romanesco) Purpose of SEM Studies ... • Research background ... SEM Workshop ap Gwynn & Richards

Transcript
Page 1: Interpretation of SEM images-iolo-printout images.pdfRomanesque Brocolli/Cauliflower (or Romanesco) Purpose of SEM Studies ... • Research background ... SEM Workshop ap Gwynn & Richards

1

“One Sees Only What One Knows”SEM Image Interpretation

Iolo ap GwynnThe University of Wales Bioimaging Laboratory

AberystwythWales

[email protected]

RMS/ESB Workshop Sorrento 2005

SEM Imaging• Information required ?

– Signal types– Resolution ?

• Specimen preparation– Preservation ?– Dehydration ?– Coating

• Microscope settings• Interpretation

– Analysis

Potential SEM Information• If used incorrectly

– Very little– Waste of time and effort

• If used to its potential– Much– Dependent upon

• Specimen preparation• Imaging conditions

• Interpretation and analysis

Quotation

“Man sieht nur was man weiss”(One sees only what one knows)

Goethe

What is this?

Romanesque Brocolli/Cauliflower (or Romanesco)

Purpose of SEM Studies

Reveal topographical surface detailSE – low voltageBSE – all voltages

Detect sub-surface informationBSE – optimise voltage

Detect compositional differencesBSE – Atomic number contrastXRA – element specific

Page 2: Interpretation of SEM images-iolo-printout images.pdfRomanesque Brocolli/Cauliflower (or Romanesco) Purpose of SEM Studies ... • Research background ... SEM Workshop ap Gwynn & Richards

2

Logical Analysis of Micrographs

CONSOLIDATED VIEW

MICROGRAPH

INTERPRETATION

A W Robards 1984

INFORMATION RETRIEVAL

Structural

CytochemicalNumerical

Instrumental effects Interactions Specimen preparation

Correlative information Supplementary considerations

Reasons for Lack of Positive Contrast e.g. BSE imaging

• Microscope settings?• Fixing/staining agent(s) has not

penetrated• Fixing/staining agent(s) has not reacted• Coating unsuitable• … etc.

Which is the correct image ?

Calcified/soft Cartilage Interface

Secondary Electron Images

Backscattered Electron Images

10 µm

8 kV 16 kV

0.5kV 1 kV 2 kV

4 kV

SE-upper detectorCalcified articular cartilage

4nm Pt/Pd

Surface of Glass Fibre Different Signal Comparison

1 kV SE4nm Pt/Pd

3 kV BSE4nm Pt/Pd

30 kV BSE4nm Pt/Pd + 60nm C

Page 3: Interpretation of SEM images-iolo-printout images.pdfRomanesque Brocolli/Cauliflower (or Romanesco) Purpose of SEM Studies ... • Research background ... SEM Workshop ap Gwynn & Richards

3

Cultured fibroblastOsmium stained

BSE images

Images courtesy of R.G.Richards

8kV

15kV

BSE

Images courtesy of R.G.Richards

Image AnalysisQuantitative dataGrey level values (256 / pixel) Measurements

length, breadth, areas etc Counting

labels

Basic Principles of Image Analysis

Areas of interest definedexpressed as binary (0 & 1) image

Measurements on binary image.

The result of ‘thresholding’

The result of ‘processing’

Page 4: Interpretation of SEM images-iolo-printout images.pdfRomanesque Brocolli/Cauliflower (or Romanesco) Purpose of SEM Studies ... • Research background ... SEM Workshop ap Gwynn & Richards

4

SomeMeasurements

Possible

1. Count the No of objects2. Area3. Length4. Breadth5. Perimeter

Others:6. Equivalent circle7. Ferets8. … and more

PC_Image

SelectMeasurements

Articular cartilagefracture propagation

= Undulation of matrix

3D Rendering by software(Alicona: Mex)

Combining Microscopies

Images courtesy of DO Meredith, AO

Take Home Messages

• Do microscopy – Carefully– Not a means to ‘decorate’ publications

• Research background– Carefully (broad sources)

• Ask a microscopist!– Before you start (potential information)– To interpret results

SEM Workshopap Gwynn & Richards 2005

ESB Sorrento


Recommended

Conventional SEM (C)SEM Environmental SEM ESEM material em paper… · Conventional SEM "(C)SEM" Environmental SEM "ESEM" 3 4 Increasing chamber pressure. 5 temperature just above
Conventional SEM (C)SEM Environmental SEM ESEM material em paper… · Conventional SEM "(C)SEM" Environmental SEM "ESEM" 3 4 Increasing chamber pressure. 5 temperature just above Documents
Sem-1 Sem-2 Sem-3 Sem-4 - University of Calcutta · 2018-06-06 · Sem ‐1 Sem‐2 Sem ... Engineering and ... Microbes in Sustainable Agriculture and Development B3. Instrumentation
Sem-1 Sem-2 Sem-3 Sem-4 - University of Calcutta · 2018-06-06 · Sem ‐1 Sem‐2 Sem ... Engineering and ... Microbes in Sustainable Agriculture and Development B3. Instrumentation Documents
Sem-1 Sem-2 Sem-3 Sem-4 - collegeadmission.in · Sem-1 Sem-2 Sem-3 Sem-4 Sem-5 Sem-6 Core Courses (CC) CC1 & 2 2Th+2P (2X4+2X2=12 Credits) CC1: Introduction to microbiology and microbial
Sem-1 Sem-2 Sem-3 Sem-4 - collegeadmission.in · Sem-1 Sem-2 Sem-3 Sem-4 Sem-5 Sem-6 Core Courses (CC) CC1 & 2 2Th+2P (2X4+2X2=12 Credits) CC1: Introduction to microbiology and microbial Documents
heritageit.edu · B.TECH / MCA TILL DATE ANY OTHER FINANCIAL ASSISTANCE APPLIED FOR / GRANTED Sem I Sem 11 Sem I Sem 11 Sem 111 Sem IV Sem 111 Sem IV Sem V Sem VI Sem V Sem VI Sem
heritageit.edu · B.TECH / MCA TILL DATE ANY OTHER FINANCIAL ASSISTANCE APPLIED FOR / GRANTED Sem I Sem 11 Sem I Sem 11 Sem 111 Sem IV Sem 111 Sem IV Sem V Sem VI Sem V Sem VI Sem Documents
Second Year-Sem III & Sem IV
Second Year-Sem III & Sem IV Documents
Rotations with Brocolli for Soilborne Disease Management ... · Pest Management Grants Final Report Contract Number: 994226 Project Title: Rotations with Broccoli for Soilborne Disease
Rotations with Brocolli for Soilborne Disease Management ... · Pest Management Grants Final Report Contract Number: 994226 Project Title: Rotations with Broccoli for Soilborne Disease Documents
Diploma IInd Sem SEM II
Diploma IInd Sem SEM II Documents
5th sem and 6th sem
5th sem and 6th sem Documents
Supporting Early Career HPE Teachers early career teachers s4.pdf · SEM 1 SEM 2 SEM 1 SEM 2 6TP2/SEM 1 SEM 2 SEM 1 6TP5/SEM 2 XNB198 Introductory Sport and Exercise Science for Teachers
Supporting Early Career HPE Teachers early career teachers s4.pdf · SEM 1 SEM 2 SEM 1 SEM 2 6TP2/SEM 1 SEM 2 SEM 1 6TP5/SEM 2 XNB198 Introductory Sport and Exercise Science for Teachers Documents
Cryo-SEM & Cryo-FIB/SEM Preparation - EMSDIASUM...Cryo-SEM & Cryo-FIB/SEM Preparation TECHNIQUES & APPLICATIONS featuring the PP3010 Cryo-SEM/Cryo-FIB/SEM Preparation System EMS_CryoSEM
Cryo-SEM & Cryo-FIB/SEM Preparation - EMSDIASUM...Cryo-SEM & Cryo-FIB/SEM Preparation TECHNIQUES & APPLICATIONS featuring the PP3010 Cryo-SEM/Cryo-FIB/SEM Preparation System EMS_CryoSEM Documents
Sem 001 sem-001
Sem 001 sem-001 Documents
Field-Emission SEM (FESEM) - MTEC · PDF fileField-Emission SEM (FESEM) Sometime called • Field Emission Gun SEM (FEG-SEM ) • High resolution SEM • Low-Voltage SEM Hitachi S8020
Field-Emission SEM (FESEM) - MTEC · PDF fileField-Emission SEM (FESEM) Sometime called • Field Emission Gun SEM (FEG-SEM ) • High resolution SEM • Low-Voltage SEM Hitachi S8020 Documents
SEM 1 SEM 4 SEM 2 SEM 3 SEM 6
SEM 1 SEM 4 SEM 2 SEM 3 SEM 6 Documents
3rd sem and 4th sem ece
3rd sem and 4th sem ece Documents
26082015_Generci Elective Sem 15 I SEM
26082015_Generci Elective Sem 15 I SEM Documents
Conseil en data stratégie - Mémorandum...Sem. 1 Sem. 2 Sem. 3 Sem. 4 Sem. 5 Sem. 6 Sem. 7 Sem. 8 Sem. 9 Sem. 10 Sem. 11 Sem. 12 Sem. 13 Sem. 14 Sem. 15 Sem. 16 Sem. 17 Sem. 18 Sem
Conseil en data stratégie - Mémorandum...Sem. 1 Sem. 2 Sem. 3 Sem. 4 Sem. 5 Sem. 6 Sem. 7 Sem. 8 Sem. 9 Sem. 10 Sem. 11 Sem. 12 Sem. 13 Sem. 14 Sem. 15 Sem. 16 Sem. 17 Sem. 18 Sem Documents
HEALTH FACTS Romanesco - Mollen FoundationRoasted Romanesco Romanesco is a cousin of Broccoli and Cauliflower. The pattern in Romanesco is a perfect example of the Fibonacci sequence
HEALTH FACTS Romanesco - Mollen FoundationRoasted Romanesco Romanesco is a cousin of Broccoli and Cauliflower. The pattern in Romanesco is a perfect example of the Fibonacci sequence Documents
𝑵𝑹𝑫 - Sernac · 0 5.000 10.000 15.000 20.000 25.000 1er sem. 2do sem. 1er sem. 2do sem. 1er sem. 2do sem. 1er sem. 2do sem. 1er sem. 2do sem. 1er sem. 2do sem. 1er sem. 2do
𝑵𝑹𝑫 - Sernac · 0 5.000 10.000 15.000 20.000 25.000 1er sem. 2do sem. 1er sem. 2do sem. 1er sem. 2do sem. 1er sem. 2do sem. 1er sem. 2do sem. 1er sem. 2do sem. 1er sem. 2do Documents