**Acknowledgements**

We are thankful to Dr. Ishaq Ahmed for inviting us to write this contribution to his edited book. We are also grateful to our research group for their help in writing up this chapter.

Furthermore, we are grateful to Dr. Kristian Molhave of DTU Nanolab, for giving us permission to adopt figures from his paper.

**79**

**Author details**

Dilawar Hassan1

Jamshoro, Pakistan

, Sidra Amin1,3, Amber Rehana Solangi1

\*Address all correspondence to: ambersolangi@gmail.com

provided the original work is properly cited.

1 National Centre of Excellence in Analytical Chemistry, University of Sindh,

2 Dr. M. A Kazi Institute of Chemistry, University of Sindh, Jamshoro, Pakistan

3 Department of Chemistry, Shaheed Benazir Bhutto University, Sindh, Pakistan

© 2019 The Author(s). Licensee IntechOpen. This chapter is distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/ by/3.0), which permits unrestricted use, distribution, and reproduction in any medium,

\* and Saima Q. Memon2

*Focused Ion Beam Tomography*

*DOI: http://dx.doi.org/10.5772/intechopen.88937*

### **Conflict of interest**

Authors have no conflict of interest.

*Focused Ion Beam Tomography DOI: http://dx.doi.org/10.5772/intechopen.88937*

*Ion Beam Techniques and Applications*

**1. Reduced osmium with potassium ferro- or ferri-cyanide**

Murphy et al. [56] Hekking et al. [58] Heymann et al. [59] Cretoiu et al. [60] Paredes-Santos et al. [61]

methyl alcohol or acetone having hydrous or anhydrous uranyl acetate and OsO4

*Variation in freeze-substitution protocols described in the literature for FIB-SEM tomography on biological* 

**2. Extended osmification with osmium-thiocarbohydrazide**

Murphy et al. [56] Leser et al. [62] Leser et al. [27] Felts et al. [63]

*Variation in chemical fixation protocols described in the literature to perform FIB-SEM tomography of* 

1. OsO4 + uranyl acetate + glutaraldehyde Murphy et al. [56] 2. OsO4 + uranyl acetate + H2O Wei et al. [57]

3. Chemical pre-fixation following by HPF/FS in OsO4 + uranyl acetate Remis et al. [69]

**3. Osmium tetroxide**

Heymann et al. [64] Murphy et al. [65] + **tannic acid** Bushby et al. [66] Armer et al. [67] Jimenez et al. [34] + **uranyl acetate** Murphy et al. [56] Leser et al. [62] Leser et al. [27] Merchán-Pérez et al. [55]

Villinger et al. [68]

A study [57] has suggested that the difference in contrast between SEM and TEM images may be because of the fixatives, additive metals, and staining agents.

It is concluded that FIB-SEM, FIB-TEM, and FIB-EDS are the latest techniques currently being used for the characterization as well as the tomographic studies of

We are thankful to Dr. Ishaq Ahmed for inviting us to write this contribution to his edited book. We are also grateful to our research group for their help in writing

Furthermore, we are grateful to Dr. Kristian Molhave of DTU Nanolab, for giv-

**78**

was used.

**Table 2.**

**Table 1.**

*biological samples.*

**5. Conclusion**

*samples cryofixed by HPF.*

materials and biological samples.

ing us permission to adopt figures from his paper.

Authors have no conflict of interest.

**Acknowledgements**

**Conflict of interest**

up this chapter.
