Electron microscopy can clarify liposome structure, if and only if done properly. The best presently available option is cryo-TEM, provided that it involves a sufficiently large samples number AND is done by a skilled practician. Lack of experience more often than not results in partially or completely misleading conclusions. Alas, the necessary combination of good equipment (incl. an adequate freezing chamber) and skill is rarely available, however.

S(mall)-A(ngle)-X(rays)-S(cattering) offers another, more easily interpretable, experimental method for vesicles characterisation, if the dispersion to be studied is, or can be made, sufficiently concentrated (>5 wght-%, better >10 wght-%). The Bragg's peaks in a SAXS diffractograme are diagnostic of multi-lamellar vesicles, whereas unilamellar vesicles or multivesicular structures (i.e., vesicles within vesicles with no parallel bilayers) yield a single diffraction peak merely. Higher order (Bragg's) peaks are observable for oligo-lamellar vesicles, with increasing intensity and visible number as the average number of (parallel) bilayers per vesicle increases.

Fluorescence (quenching) has also been advocated for vesicle lamellarity studies. Using this, or some other spectroscopy (e.g., NMR signal quenching) to similar end requires VERY careful checks and calibrations for gaining reliable results.

Since the question of liposome lamellarity is most often asked in the context of formulation / delivery work, a vesicles content release study may be researchers' best choice. A CAREFULLY EXECUTED and WELL MODELLED such study can namely reveal not only how much of the intended payload is encapsulated into vesicles and/or associated with the vesicles bilayers (resulting in two different release kinetics), but also uncover the AVERAGE number of bilayers ("lamellae") per vesicle: the release rate of the same encapsulated / associated content from 2-, 3-, 4-'lamellars' vesicles is 1/2, 1/3, 1/4 of the release rate from the corresponding unilamellar vesicles, since the cargo then has to cross 2, 3, 4 barriers in a row. Detailed modelling (considering at least the rates of cargo crossing AND SEPARATELY release from EACH involved bilayer) is a key to the resulting data proper analysis. Its basis is the reference data set obtained with properly equilibrated unilamellar vesicles (not smaller than 70 nm for the stiff bilayers and 50 nm for the fluid-chains bilayers to avoid stress induced, release accelerating defects), which affords said rates values for single bilayers.

In order to a proper morphology study for liposome a cryo-SEM and cryo-TEM imaging should be performed. Your concern of drying is taken care in cryo imaging as it is done in the native state itself. I am not sure why you expect to observe lamellarity in SEM images (I expect those in TEM images).

Methods of measuring lamellarity is mentioned in this paper Article Parameters influencing the determination of liposome lamella...

Electron microscopy can clarify liposome structure, if and only if done properly. The best presently available option is cryo-TEM, provided that it involves a sufficiently large samples number AND is done by a skilled practician. Lack of experience more often than not results in partially or completely misleading conclusions. Alas, the necessary combination of good equipment (incl. an adequate freezing chamber) and skill is rarely available, however.

S(mall)-A(ngle)-X(rays)-S(cattering) offers another, more easily interpretable, experimental method for vesicles characterisation, if the dispersion to be studied is, or can be made, sufficiently concentrated (>5 wght-%, better >10 wght-%). The Bragg's peaks in a SAXS diffractograme are diagnostic of multi-lamellar vesicles, whereas unilamellar vesicles or multivesicular structures (i.e., vesicles within vesicles with no parallel bilayers) yield a single diffraction peak merely. Higher order (Bragg's) peaks are observable for oligo-lamellar vesicles, with increasing intensity and visible number as the average number of (parallel) bilayers per vesicle increases.

Fluorescence (quenching) has also been advocated for vesicle lamellarity studies. Using this, or some other spectroscopy (e.g., NMR signal quenching) to similar end requires VERY careful checks and calibrations for gaining reliable results.

Since the question of liposome lamellarity is most often asked in the context of formulation / delivery work, a vesicles content release study may be researchers' best choice. A CAREFULLY EXECUTED and WELL MODELLED such study can namely reveal not only how much of the intended payload is encapsulated into vesicles and/or associated with the vesicles bilayers (resulting in two different release kinetics), but also uncover the AVERAGE number of bilayers ("lamellae") per vesicle: the release rate of the same encapsulated / associated content from 2-, 3-, 4-'lamellars' vesicles is 1/2, 1/3, 1/4 of the release rate from the corresponding unilamellar vesicles, since the cargo then has to cross 2, 3, 4 barriers in a row. Detailed modelling (considering at least the rates of cargo crossing AND SEPARATELY release from EACH involved bilayer) is a key to the resulting data proper analysis. Its basis is the reference data set obtained with properly equilibrated unilamellar vesicles (not smaller than 70 nm for the stiff bilayers and 50 nm for the fluid-chains bilayers to avoid stress induced, release accelerating defects), which affords said rates values for single bilayers.

Thanks everyone for giving such invaluable suggestions, it'll definitely help me to ease up my work.

You can use confocal microscopy. When you encapsulate a molecule that is fluorescent and also hydrophobic, you can see, if the liposome is big enough, in the confocal microscopy, the lamellae. Search for: confocal microscopy liposome rhodamine